Hepatitis C virus inhibitors

ABSTRACT

This disclosure concerns novel compounds of Formula (I) as defined in the specification and compositions comprising such novel compounds. These compounds are useful antiviral agents, especially in inhibiting the function of the NS5A protein encoded by Hepatitis C virus (HCV). Thus, the disclosure also concerns a method of treating HCV related diseases or conditions by use of these novel compounds or a composition comprising such novel compounds.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. Non-Provisional applicationSer. No. 12/731,935 filed Mar. 25, 2010 which claims the benefit of U.S.Provisional Application Ser. No. 61/164,579 filed Mar. 30, 2009.

FIELD OF THE INVENTION

The present disclosure is generally directed to antiviral compounds, andmore specifically directed to compounds which can inhibit the functionof the NS5A protein encoded by Hepatitis C virus (HCV), compositionscomprising such compounds, and methods for inhibiting the function ofthe NS5A protein.

BACKGROUND OF THE INVENTION

HCV is a major human pathogen, infecting an estimated 170 millionpersons worldwide—roughly five times the number infected by humanimmunodeficiency virus type 1. A substantial fraction of these HCVinfected individuals develop serious progressive liver disease,including cirrhosis and hepatocellular carcinoma.

The current standard of care for HCV, which employs a combination ofpegylated-interferon and ribavirin, has a non-optimal success rate inachieving sustained viral response and causes numerous side effects.Thus, there is a clear and long-felt need to develop effective therapiesto address this undermet medical need.

HCV is a positive-stranded RNA virus. Based on a comparison of thededuced amino acid sequence and the extensive similarity in the 5′untranslated region, HCV has been classified as a separate genus in theFlaviviridae family. All members of the Flaviviridae family haveenveloped virions that contain a positive stranded RNA genome encodingall known virus-specific proteins via translation of a single,uninterrupted, open reading frame.

Considerable heterogeneity is found within the nucleotide and encodedamino acid sequence throughout the HCV genome due to the high error rateof the encoded RNA dependent RNA polymerase which lacks a proof-readingcapability. At least six major genotypes have been characterized, andmore than 50 subtypes have been described with distribution worldwide.The clinical significance of the genetic heterogeneity of HCV hasdemonstrated a propensity for mutations to arise during monotherapytreatment, thus additional treatment options for use are desired. Thepossible modulator effect of genotypes on pathogenesis and therapyremains elusive.

The single strand HCV RNA genome is approximately 9500 nucleotides inlength and has a single open reading frame (ORF) encoding a single largepolyprotein of about 3000 amino acids. In infected cells, thispolyprotein is cleaved at multiple sites by cellular and viral proteasesto produce the structural and non-structural (NS) proteins. In the caseof HCV, the generation of mature non-structural proteins (NS2, NS3,NS4A, NS4B, NS5A, and NS5B) is effected by two viral proteases. Thefirst one is believed to be a metalloprotease and cleaves at the NS2-NS3junction; the second one is a serine protease contained within theN-terminal region of NS3 (also referred to herein as NS3 protease) andmediates all the subsequent cleavages downstream of NS3, both in cis, atthe NS3-NS4A cleavage site, and in trans, for the remaining NS4A-NS4B,NS4B-NS5A, NS5A-NS5B sites. The NS4A protein appears to serve multiplefunctions by both acting as a cofactor for the NS3 protease andassisting in the membrane localization of NS3 and other viral replicasecomponents. The formation of a NS3-NS4A complex is necessary for properprotease activity resulting in increased proteolytic efficiency of thecleavage events. The NS3 protein also exhibits nucleoside triphosphataseand RNA helicase activities. NS5B (also referred to herein as HCVpolymerase) is a RNA-dependent RNA polymerase that is involved in thereplication of HCV with other HCV proteins, including NS5A, in areplicase complex.

Compounds useful for treating HCV-infected patients are desired whichselectively inhibit HCV viral replication. In particular, compoundswhich are effective to inhibit the function of the NS5A protein aredesired. The HCV NS5A protein is described, for example, in thefollowing references: S. L. Tan, et al., Virology, 284:1-12 (2001);K.-J. Park, et al., J. Biol. Chem., 30711-30718 (2003); T. L.Tellinghuisen, et al., Nature, 435, 374 (2005); R. A. Love, et al., J.Virol, 83, 4395 (2009); N. Appel, et al., J. Biol. Chem., 281, 9833(2006); L. Huang, J. Biol. Chem., 280, 36417 (2005); C. Rice, et al.,WO2006093867.

SUMMARY OF THE INVENTION

The present invention provides compounds which selectively inhibit HCVviral replication, as characterized by Formula (I):

or a pharmaceutically acceptable salt thereof, wherein:

-   -   s is 0 or 1;    -   L is -L¹-L²-, wherein L¹ and L² are independently selected from:

-   -    provided that at least one of L¹ and L² is other than

-   -   Y and Y′ are independently oxygen (O) or NH;    -   R¹ is hydrogen or —C(O)R^(x);    -   R² is hydrogen or —C(O)R^(y);    -   R^(x) and R^(y) are independently selected from cycloalkyl,        heteroaryl, heterocyclyl, alkoxy, and alkyl substituted with one        or more substituents independently selected from aryl, alkenyl,        cycloalkyl, heterocyclyl, heteroaryl, —OR³,    -   —C(O)OR⁴, —NR^(a)R^(b), and —C(O)NR^(c)R^(d),    -   wherein aryl and heteroaryl may optionally be substituted with        one or more substituents independently selected from alkyl,        haloalkyl, arylalkyl, heterocyclyl, heterocyclylalkyl, halogen,        cyano, nitro, —C(O)OR⁴, OR⁵, —NR^(a)R^(b), (NR^(a)R^(b))alkyl,        and (MeO)(HO)P(O)O—, and    -   wherein cycloalkyl and heterocyclyl may optionally be fused onto        an aromatic ring and may optionally be substituted with one or        more substituents independently selected from alkyl, hydroxyl,        halogen, aryl, —NR^(a)R^(b), oxo, and —C(O)OR⁴;    -   R³ is hydrogen, alkyl, or arylalkyl;    -   R⁴ is alkyl or arylalkyl;    -   R⁵ is hydrogen, alkyl, or arylalkyl;    -   R^(a) and R^(b) are independently selected from hydrogen, alkyl,        cycloalkyl, arylalkyl, heteroaryl, —C(O)R⁶, —C(O)OR⁷,        —C(O)NR^(c)R^(d), and (NR^(c)R^(d))alkyl, or alternatively,        R^(a) and R^(b), together with the nitrogen atom to which they        are attached, form a five- or six-membered ring or bridged        bicyclic ring structure, wherein said five- or six-membered ring        or bridged bicyclic ring structure optionally may contain one or        two additional heteroatoms independently selected from nitrogen,        oxygen, and sulfur and may contain one, two, or three        substituents independently selected from C₁ to C₆ alkyl, C₁ to        C₄ haloalkyl, aryl, hydroxyl, C₁ to C₆ alkoxy, C₁ to C₄        haloalkoxy, and halogen;    -   R⁶ is alkyl;    -   R⁷ is alkyl, arylalkyl, cycloalkyl, or haloalkyl;    -   R¹⁰⁰ and R¹¹⁰ are independently selected from hydrogen, alkyl,        cyanoalkyl, and halo;    -   R^(c) and R^(d) are independently selected from hydrogen, alkyl,        arylalkyl, and cycloalkyl.

The compounds of the present disclosure can be effective to inhibit thefunction of the HCV NS5A protein. In particular, the compounds of thepresent disclosure can be effective to inhibit the HCV 1b genotype ormultiple genotypes of HCV. Therefore, this invention also encompasses:(1) compositions comprising a compound of Formula (I), or apharmaceutically acceptable salt thereof, and a pharmaceuticallyacceptable carrier; and (2) a method of treating an HCV infection in apatient, comprising administering to the patient a therapeuticallyeffective amount of a compound of Formula (I), or a pharmaceuticallyacceptable salt thereof.

DETAILED DESCRIPTION OF THE INVENTION

In a first aspect of the present disclosure compounds of Formula (I) areprovided:

In a first embodiment of the first aspect, the present disclosureprovides a compound of Formula (I), or a pharmaceutically acceptablesalt or solvate thereof, wherein:

-   -   s is 0 or 1;    -   L is -L¹-L²-, wherein L¹ and L² are independently selected from:

-   -    provided that at least one of L¹ and L² is other than

-   -   Y and Y′ are independently oxygen (O) or NH;    -   R¹ is hydrogen or —C(O)R^(x);    -   R² is hydrogen or —C(O)R^(y);    -   R^(x) and R^(y) are independently selected from cycloalkyl,        heteroaryl, heterocyclyl, alkoxy, and alkyl substituted with one        or more substituents independently selected from aryl, alkenyl,        cycloalkyl, heterocyclyl, heteroaryl, —OR³,    -   —C(O)OR⁴, —NR^(a)R^(b), and —C(O)NR^(c)R^(d),    -   wherein aryl and heteroaryl may optionally be substituted with        one or more substituents independently selected from alkyl,        haloalkyl, arylalkyl, heterocyclyl, heterocyclylalkyl, halogen,        cyano, nitro, —C(O)OR⁴, OR⁵, —NR^(a)R^(b), (NR^(a)R^(b))alkyl,        and (MeO)(HO)P(O)O—, and    -   wherein cycloalkyl and heterocyclyl may optionally be fused onto        an aromatic ring and may optionally be substituted with one or        more substituents independently selected from alkyl, hydroxyl,        halogen, aryl, —NR^(a)R^(b), oxo, and —C(O)OR⁴;    -   R³ is hydrogen, alkyl, or arylalkyl;    -   R⁴ is alkyl or arylalkyl;    -   R⁵ is hydrogen, alkyl, or arylalkyl;    -   R^(a) and R^(b) are independently selected from hydrogen, alkyl,        cycloalkyl, arylalkyl, heteroaryl, —C(O)R⁶, —C(O)OR⁷,        —C(O)NR^(c)R^(d), and (NR^(c)R^(d))alkyl, or alternatively,        R^(a) and R^(b), together with the nitrogen atom to which they        are attached, form a five- or six-membered ring or bridged        bicyclic ring structure, wherein said five- or six-membered ring        or bridged bicyclic ring structure optionally may contain one or        two additional heteroatoms independently selected from nitrogen,        oxygen, and sulfur and may contain one, two, or three        substituents independently selected from C₁ to C₆ alkyl, C₁ to        C₄ haloalkyl, aryl, hydroxyl, C₁ to C₆ alkoxy, C₁ to C₄        haloalkoxy, and halogen;    -   R⁶ is alkyl;    -   R⁷ is alkyl, arylalkyl, or haloalkyl;    -   R¹⁰⁰ and R¹¹⁰ are independently selected from hydrogen, alkyl,        cyanoalkyl, and halo;    -   R^(c) and R^(d) are independently selected from hydrogen, alkyl,        arylalkyl, and cycloalkyl.

In a first class of compounds of the first embodiment, the presentdisclosure provides a compound of Formula (I), or a pharmaceuticallyacceptable salt or solvate thereof, wherein s is 0.

In a second class of compounds of the first embodiment, the presentdisclosure provides a compound of Formula (I), or a pharmaceuticallyacceptable salt or solvate thereof, wherein s is 1.

In a second embodiment of the first aspect, the present disclosureprovides a compound of Formula (I), or a pharmaceutically acceptablesalt or solvate thereof, wherein:

-   -   s is 0 or 1;    -   L is selected from:

-   -   Y and Y′ are independently oxygen (O) or NH;    -   R¹ is hydrogen or —C(O)R^(x);    -   R² is hydrogen or —C(O)R^(y);    -   R^(x) and R^(y) are independently selected from cycloalkyl,        heteroaryl, heterocyclyl, alkoxy, and alkyl substituted with one        or more substituents independently selected from aryl, alkenyl,        cycloalkyl, heterocyclyl, heteroaryl, —OR³,    -   —C(O)OR⁴, —NR^(a)R^(b), and —C(O)NR^(c)R^(d),    -   wherein aryl and heteroaryl may optionally be substituted with        one or more substituents independently selected from alkyl,        haloalkyl, arylalkyl, heterocyclyl, heterocyclylalkyl, halogen,        cyano, nitro, —C(O)OR⁴, OR⁵, —NR^(a)R^(b), (NR^(a)R^(b))alkyl,        and (MeO)(HO)P(O)O—, and    -   wherein cycloalkyl and heterocyclyl may optionally be fused onto        an aromatic ring and may optionally be substituted with one or        more substituents independently selected from alkyl, hydroxyl,        halogen, aryl, —NR^(a)R^(b), oxo, and —C(O)OR⁴;    -   R³ is hydrogen, alkyl, or arylalkyl;    -   R⁴ is alkyl or arylalkyl;    -   R⁵ is hydrogen, alkyl, or arylalkyl;    -   R^(a) and R^(b) are independently selected from hydrogen, alkyl,        cycloalkyl, arylalkyl, heteroaryl, —C(O)R⁶, —C(O)OR⁷,        —C(O)NR^(c)R^(d), and (NR^(c)R^(d))alkyl, or alternatively,        R^(a) and R^(b), together with the nitrogen atom to which they        are attached, form a five- or six-membered ring or bridged        bicyclic ring structure, wherein said five- or six-membered ring        or bridged bicyclic ring structure optionally may contain one or        two additional heteroatoms independently selected from nitrogen,        oxygen, and sulfur and may contain one, two, or three        substituents independently selected from C₁ to C₆ alkyl, C₁ to        C₄ haloalkyl, aryl, hydroxyl, C₁ to C₆ alkoxy, C₁ to C₄        haloalkoxy, and halogen;    -   R⁶ is alkyl;    -   R⁷ is alkyl, arylalkyl, or haloalkyl;    -   R¹⁰⁰ and R¹¹⁰ are independently selected from hydrogen, alkyl,        cyanoalkyl, and halo;    -   R^(c) and R^(d) are independently selected from hydrogen, alkyl,        arylalkyl, and cycloalkyl.

In a first class of compounds in the second embodiment, the presentdisclosure provides a compound of Formula (I), or a pharmaceuticallyacceptable salt or solvate thereof, wherein L is

In a second class of compounds of the second embodiment, the presentdisclosure provides a compound of Formula (I), or a pharmaceuticallyacceptable salt or solvate thereof, wherein L is

In a third class of compounds of the second embodiment, the presentdisclosure provides a compound of Formula (I), or a pharmaceuticallyacceptable salt or solvate thereof, wherein L is

In a fourth class of compounds of the second embodiment, the presentdisclosure provides a compound of Formula (I), or a pharmaceuticallyacceptable salt or solvate thereof, wherein L is

In a fifth class of compounds of the second embodiment, the presentdisclosure provides a compound of Formula (I), or a pharmaceuticallyacceptable salt or solvate thereof, wherein L is

In a sixth class of compounds of the second embodiment, the presentdisclosure provides a compound of Formula (I), or a pharmaceuticallyacceptable salt or solvate thereof, wherein L is

In a seventh class of compounds of the second embodiment, the presentdisclosure provides a compound of Formula (I), or a pharmaceuticallyacceptable salt or solvate thereof, wherein L is

In an eighth class of compounds of the second embodiment, the presentdisclosure provides a compound of Formula (I), or a pharmaceuticallyacceptable salt or solvate thereof, wherein L is

In a ninth class of compounds of the second embodiment, the presentdisclosure provides a compound of Formula (I), or a pharmaceuticallyacceptable salt or solvate thereof, wherein L is

In a third embodiment of the first aspect, the present disclosureprovides a compound of Formula (I), or a pharmaceutically acceptablesalt or solvate thereof, wherein:

-   -   s is 0 or 1;    -   L is -L¹-L²-, wherein L¹ and L² are independently selected from:

-   -    provided that at least one of L¹ and L² is other than

-   -   Y and Y′ are each NH;    -   R¹ is hydrogen or —C(O)R^(x);    -   R² is hydrogen or —C(O)R^(y);    -   R^(x) and R^(y) are independently selected from cycloalkyl,        heteroaryl, heterocyclyl, alkoxy, and alkyl substituted with one        or more substituents independently selected from aryl, alkenyl,        cycloalkyl, heterocyclyl, heteroaryl, —OR³,    -   —C(O)OR⁴, —NR^(a)R^(b), and —C(O)NR^(c)R^(d),    -   wherein aryl and heteroaryl may optionally be substituted with        one or more substituents independently selected from alkyl,        haloalkyl, arylalkyl, heterocyclyl, heterocyclylalkyl, halogen,        cyano, nitro, —C(O)OR⁴, OR⁵, —NR^(a)R^(b), (NR^(a)R^(b))alkyl,        and (MeO)(HO)P(O)O—, and    -   wherein cycloalkyl and heterocyclyl may optionally be fused onto        an aromatic ring and may optionally be substituted with one or        more substituents independently selected from alkyl, hydroxyl,        halogen, aryl, —NR^(a)R^(b), oxo, and —C(O)OR⁴;    -   R³ is hydrogen, alkyl, or arylalkyl;    -   R⁴ is alkyl or arylalkyl;    -   R⁵ is hydrogen, alkyl, or arylalkyl;    -   R^(a) and R^(b) are independently selected from hydrogen, alkyl,        cycloalkyl, arylalkyl, heteroaryl, —C(O)R⁶, —C(O)OR⁷,        —C(O)NR^(c)R^(d), and (NR^(c)R^(d))alkyl, or alternatively,        R^(a) and R^(b), together with the nitrogen atom to which they        are attached, form a five- or six-membered ring or bridged        bicyclic ring structure, wherein said five- or six-membered ring        or bridged bicyclic ring structure optionally may contain one or        two additional heteroatoms independently selected from nitrogen,        oxygen, and sulfur and may contain one, two, or three        substituents independently selected from C₁ to C₆ alkyl, C₁ to        C₄ haloalkyl, aryl, hydroxyl, C₁ to C₆ alkoxy, C₁ to C₄        haloalkoxy, and halogen;    -   R⁶ is alkyl;    -   R² is alkyl, arylalkyl, or haloalkyl;    -   R¹⁰⁰ and R¹¹⁰ are independently selected from hydrogen, alkyl,        cyanoalkyl, and halo;    -   R^(c) and R^(d) are independently selected from hydrogen, alkyl,        arylalkyl, and cycloalkyl.

In a fourth embodiment of the first aspect, the present disclosureprovides a compound of Formula (I), or a pharmaceutically acceptablesalt or solvate thereof, wherein:

-   -   s is 0 or 1;    -   L is -L¹-L²-, wherein L¹ and L² are independently selected from:

-   -    provided that at least one of L¹ and L² is other than

-   -   Y is oxygen (O), and Y′ is NH;    -   R¹ is hydrogen or —C(O)R^(x);    -   R² is hydrogen or —C(O)R^(y);    -   R^(x) and R^(y) are independently selected from cycloalkyl,        heteroaryl, heterocyclyl, alkoxy, and alkyl substituted with one        or more substituents independently selected from aryl, alkenyl,        cycloalkyl, heterocyclyl, heteroaryl, —OR³,    -   —C(O)OR⁴, —NR^(a)R^(b), and —C(O)NR^(c)R^(d),    -   wherein aryl and heteroaryl may optionally be substituted with        one or more substituents independently selected from alkyl,        haloalkyl, arylalkyl, heterocyclyl, heterocyclylalkyl, halogen,        cyano, nitro, —C(O)OR⁴, OR⁵, —NR^(a)R^(b), (NR^(a)R^(b))alkyl,        and (MeO)(HO)P(O)O—, and    -   wherein cycloalkyl and heterocyclyl may optionally be fused onto        an aromatic ring and may optionally be substituted with one or        more substituents independently selected from alkyl, hydroxyl,        halogen, aryl, —NR^(a)R^(b), oxo, and —C(O)OR⁴;    -   R³ is hydrogen, alkyl, or arylalkyl;    -   R⁴ is alkyl or arylalkyl;    -   R⁵ is hydrogen, alkyl, or arylalkyl;    -   R^(a) and R^(b) are independently selected from hydrogen, alkyl,        cycloalkyl, arylalkyl, heteroaryl, —C(O)R⁶, —C(O)OR⁷,        —C(O)NR^(c)R^(d), and (NR^(c)R^(d))alkyl, or alternatively,        R^(a) and R^(b), together with the nitrogen atom to which they        are attached, form a five- or six-membered ring or bridged        bicyclic ring structure, wherein said five- or six-membered ring        or bridged bicyclic ring structure optionally may contain one or        two additional heteroatoms independently selected from nitrogen,        oxygen, and sulfur and may contain one, two, or three        substituents independently selected from C₁ to C₆ alkyl, C₁ to        C₄ haloalkyl, aryl, hydroxyl, C₁ to C₆ alkoxy, C₁ to C₄        haloalkoxy, and halogen;    -   R⁶ is alkyl;    -   R⁷ is alkyl, arylalkyl, or haloalkyl;    -   R¹⁰⁰ and R¹¹⁰ are independently selected from hydrogen, alkyl,        cyanoalkyl, and halo;    -   R^(c) and R^(d) are independently selected from hydrogen, alkyl,        arylalkyl, and cycloalkyl.

In a fifth embodiment of the first aspect, the present disclosureprovides a compound of Formula (I), or a pharmaceutically acceptablesalt or solvate thereof, wherein:

-   -   s is 0 or 1;    -   L is -L¹-L²-, wherein L¹ and L² are independently selected from:

-   -    provided that at least one of L¹ and L² is other than

-   -   R¹ is —C(O)R^(x);    -   R² is —C(O)R^(y);    -   R^(x) and R^(y) are independently alkyl substituted by at least        one —NR^(a)R^(b), characterized by Formula (A):

wherein:

-   -   m is 0 or 1;    -   R⁸ is hydrogen or alkyl;    -   R⁹ is selected from hydrogen, cycloalkyl, aryl, heteroaryl,        heterocyclyl, and alkyl optionally substituted with a        substituent selected from aryl, alkenyl, cycloalkyl,        heterocyclyl, heteroaryl, heterobicyclyl, —OR³, —C(O)OR⁴,        —NR^(a)R^(b), and —C(O)NR^(c)R^(d),    -   wherein aryl and heteroaryl may optionally be substituted with        one or more substituents independently selected from alkyl,        haloalkyl, arylalkyl, heterocyclyl, heterocyclylalkyl, halogen,        cyano, nitro, —C(O)OR⁴, OR⁵, —NR^(a)R^(b), (NR^(a)R^(b))alkyl,        and (MeO)(HO)P(O)O—, and    -   wherein cycloalkyl and heterocyclyl may optionally be fused onto        an aromatic ring and may optionally be substituted with one or        more substituents independently selected from alkyl, hydroxyl,        halogen, aryl, —NR^(a)R^(b), oxo, and —C(O)OR⁴;    -   R¹⁰⁰ and R¹¹⁰ are independently selected from hydrogen and halo;    -   R³ is hydrogen, alkyl, or arylalkyl;    -   R⁴ is alkyl or arylalkyl;    -   R⁵ is hydrogen, alkyl, or arylalkyl;    -   R^(a) and R^(b) are independently selected from hydrogen, alkyl,        cycloalkyl, arylalkyl, heteroaryl, —C(O)R⁶, —C(O)OR⁷,        —C(O)NR^(c)R^(d), and (NR^(c)R^(d))alkyl, or alternatively,        R^(a) and R^(b), together with the nitrogen atom to which they        are attached, form a five- or six-membered ring or bridged        bicyclic ring structure, wherein said five- or six-membered ring        or bridged bicyclic ring structure optionally may contain one or        two additional heteroatoms independently selected from nitrogen,        oxygen, and sulfur and may contain one, two, or three        substituents independently selected from C₁ to C₆ alkyl, C₁ to        C₄ haloalkyl, aryl, hydroxyl, C₁ to C₆ alkoxy, C₁ to C₄        haloalkoxy, and halogen;    -   R⁶ is alkyl;    -   R² is alkyl, arylalkyl, or haloalkyl;    -   R¹⁰⁰ and R¹¹⁰ are independently selected from hydrogen, alkyl,        cyanoalkyl, and halo; and    -   R^(c) and R^(d) are independently selected from hydrogen, alkyl,        arylalkyl, and cycloalkyl.

In a sixth embodiment of the first aspect, the present disclosureprovides a compound of Formula (I), or a pharmaceutically acceptablesalt or solvate thereof, wherein:

-   -   s is 0 or 1;    -   L is -L¹-L²-, wherein L¹ and L² are independently selected from:

-   -    provided that at least one of L¹ and L² is other than

-   -   R¹ is —C(O)R^(x);    -   R² is —C(O)R^(y);    -   R^(x) and R^(y) are independently alkyl substituted by at least        one —NR^(a)R^(b), characterized by Formula (A):

wherein:

-   -   m is 0;    -   R⁸ is hydrogen or C₁ to C₄ alkyl;    -   R⁹ is selected from hydrogen, C₁ to C₆ alkyl optionally        substituted with —OR¹², C₃ to C₆ cycloalkyl, allyl,        —CH₂C(O)NR^(c)R^(d), (NR^(c)R^(d))alkyl,

-   -   wherein j is 0 or 1;    -   k is 1, 2, or 3;    -   n is 0 or an integer selected from 1 through 4;    -   each R¹⁰ is independently hydrogen, C₁ to C₄ alkyl, C₁ to C₄        haloalkyl, halogen, nitro, —OBn, or (MeO)(OH)P(O)O—;    -   R¹¹ is hydrogen, C₁ to C₄ alkyl, or benzyl;    -   R¹² is hydrogen, C₁ to C₄ alkyl, or benzyl;    -   R^(a) is hydrogen or C₁ to C₄ alkyl;    -   R^(b) is C₁ to C₄ alkyl, C₃ to C₆ cycloalkyl, benzyl, 3-pyridyl,        pyrimidin-5-yl, acetyl, —C(O)OR⁷, or —C(O)NR^(c)R^(d);    -   R⁷ is C₁ to C₄ alkyl or C₁ to C₄ haloalkyl;    -   R¹⁰⁰ and R¹¹⁰ are independently selected from hydrogen and halo;        and    -   R^(c) is hydrogen or C₁ to C₄ alkyl; and    -   R^(d) is hydrogen, C₁ to C₄ alkyl, or C₃ to C₆ cycloalkyl.

In a seventh embodiment of the first aspect, the present disclosureprovides a compound of Formula (I), or a pharmaceutically acceptablesalt or solvate thereof, wherein:

-   -   s is 0 or 1;    -   L is -L¹-L²-, wherein L¹ and L² are independently selected from:

-   -    provided that at least one of L¹ and L² is other than

-   -   R¹ is —C(O)R^(x);    -   R² is —C(O)R^(y);    -   R^(x) and R^(y) are independently alkyl substituted by at least        one —NR^(a)R^(b), characterized by Formula (A):

wherein:

-   -   m is 0;    -   R⁸ is hydrogen;    -   R⁹ is phenyl optionally substituted with one up to five        substituents independently selected from C₁ to C₆ alkyl, C₁ to        C₄ haloalkyl, halogen, C₁ to C₆ alkoxy, hydroxyl, cyano, and        nitro; and    -   NR^(a)R^(b) is a heterocyclyl or heterobicyclyl group selected        from:

-   -   wherein n is 0, 1, or 2;    -   each R¹³ is independently selected from C₁ to C₆ alkyl, phenyl,        trifluoromethyl, halogen, hydroxyl, methoxy, and oxo;    -   R¹⁴ is C₁ to C₆ alkyl, phenyl, benzyl, or C(O)OR¹⁵ group,        wherein R¹⁵ is C₁ to C₄ alkyl, phenyl, or benzyl; and    -   R¹⁰⁰ and R¹¹⁰ are independently selected from hydrogen and halo.

In an eighth embodiment of the first aspect, the present disclosureprovides a compound of Formula (I), or a pharmaceutically acceptablesalt or solvate thereof, wherein:

-   -   s is 0 or 1;    -   L is -L¹-L²-, wherein L¹ and L² are independently selected from:

-   -    provided that at least one of L¹ and L² is other than

-   -   R¹ is —C(O)R^(x);    -   R² is —C(O)R^(y);    -   R^(x) and R^(y) are independently alkyl substituted by at least        one —NR^(a)R^(b), characterized by Formula (A):

wherein:

-   -   m is 1;    -   R⁸ is hydrogen;    -   R⁹ is C₁ to C₆ alkyl, arylalkyl, or heteroarylalkyl;    -   R¹⁰⁰ and R¹¹⁰ are independently selected from hydrogen and halo;    -   R^(a) is hydrogen; and    -   R^(b) is —C(O)OR⁷, wherein R⁷ is C₁ to C₆ alkyl.

In a ninth embodiment of the first aspect, the present disclosureprovides a compound of Formula (I), or a pharmaceutically acceptablesalt or solvate thereof, wherein:

-   -   s is 0 or 1;    -   L is -L¹-L²-, wherein L¹ and L² are independently selected from:

-   -    provided that at least one of L¹ and L² is other than

-   -   R¹ is —C(O)R^(x);    -   R² is —C(O)R^(y);    -   R^(x) and R^(y) are heteroaryl or heterocyclyl independently        selected from:

-   -   wherein n is 0 or an integer selected from 1 through 4;    -   each R¹³ is independently selected from hydrogen, C₁ to C₆        alkyl, C₁ to C₄ haloalkyl, phenyl, benzyl, C₁ to C₆ alkoxy, C₁        to C₄ haloalkoxy, heterocyclyl, halogen, —NR^(c)R^(d), hydroxyl,        cyano, and oxo, where R^(c) and R^(d) are independently hydrogen        or C₁ to C₄ alkyl; and    -   R¹⁴ is hydrogen (H), C₁ to C₆ alkyl, benzyl, or —C(O)OR⁴,        wherein R⁴ is C₁ to C₆ alkyl.

In a tenth embodiment of the first aspect, the present disclosureprovides a compound of Formula (I), or a pharmaceutically acceptablesalt or solvate thereof, wherein:

-   -   s is 0 or 1;    -   L is -L¹-L²-, wherein L¹ and L² are independently selected from:

-   -    provided that at least one of L¹ and L² is other than

-   -   R¹ is —C(O)R^(x);    -   R² is —C(O)R^(y);    -   R^(x) and R^(y) are cycloalkyl independently selected from:

-   -   wherein j is 0, 1, 2, or 3;    -   k is 0, 1, or 2;    -   n is 0 or an integer selected from 1 through 4;    -   each R¹³ is independently selected from hydrogen, C₁ to C₆        alkyl, C₁ to C₄ haloalkyl, C₁ to C₆ alkoxy, halogen, hydroxyl,        cyano, and nitro; and    -   R^(a) and R^(b) are each independently hydrogen, C₁ to C₆ alkyl,        or C(O)OR⁷,    -   wherein R⁷ is C₁ to C₆ alkyl.

In an eleventh embodiment of the first aspect, the present disclosureprovides a compound of Formula (I), or a pharmaceutically acceptablesalt or solvate thereof, wherein:

-   -   s is 0 or 1;    -   L is -L¹-L²-, wherein L¹ and L² are independently selected from:

-   -    provided that at least one of L¹ and L² is other than

-   -   R¹ is —C(O)R^(x);    -   R² is —C(O)R^(y);    -   R^(x) and R^(y) are independently arylalkyl, wherein aryl part        of said arylalkyl may optionally be substituted with        (NR^(a)R^(b))alkyl; and    -   R^(a) and R^(b) are independently hydrogen, C₁ to C₆ alkyl, or        benzyl, or alternatively, R^(a) and R^(b), together with the        nitrogen atom to which they are attached, form a five- or        six-membered ring selected from

-   -    wherein R¹⁵ is hydrogen,    -   C₁ to C₆ alkyl, or benzyl.

In a twelfth embodiment of the first aspect, the present disclosureprovides a compound of Formula (I), or a pharmaceutically acceptablesalt or solvate thereof, wherein:

-   -   s is 0 or 1;    -   L is -L¹-L²-, wherein L¹ and L² are independently selected from:

-   -    provided that at least one of L¹ and L² is other than

-   -   R¹ is —C(O)R^(x);    -   R² is —C(O)R^(y); and    -   R^(x) and R^(y) are the same and are selected from the group        consisting of:

wherein a squiggle bond (

) in the structure indicates that a stereogenic center to which the bondis attached can take either (R)- or (S)-configuration so long aschemical bonding principles are not violated.

In a thirteenth embodiment of the first aspect, the present disclosureprovides a compound of Formula (I), or a pharmaceutically acceptablesalt or solvate thereof, wherein:

-   -   S is 0 or 1;    -   R¹ is —C(O)R^(x);    -   R² is —C(O)R^(y); and    -   R^(x) and R^(y) are both t-butoxy.

In a fourteenth embodiment of the first aspect, the present disclosureprovides a compound of Formula (I), or a pharmaceutically acceptablesalt or solvate thereof, wherein:

-   -   s is 0 or 1; and    -   R¹ and R² are both hydrogen.

In a fiftheenth embodiment of the first aspect, the present disclosureprovides a compound of Formula (II):

or a pharmaceutically acceptable salt thereof, wherein:

-   -   s is 0 or 1;    -   L is -L¹-L²-, wherein L¹ and L² are independently selected from:

-   -    provided that at least one of L¹ and L² is other than

-   -   Y and Y′ are independently oxygen (O) or NH;    -   R¹ is hydrogen or —C(O)R^(x);    -   R² is hydrogen or —C(O)R^(y);    -   R^(x) and R^(y) are independently selected from cycloalkyl,        heteroaryl, heterocyclyl, alkoxy, and alkyl substituted with one        or more substituents independently selected from aryl, alkenyl,        cycloalkyl, heterocyclyl, heteroaryl, —OR³,    -   —C(O)OR⁴, —NR^(a)R^(b), and —C(O)NR^(c)R^(d),    -   wherein aryl and heteroaryl may optionally be substituted with        one or more substituents independently selected from alkyl,        haloalkyl, arylalkyl, heterocyclyl, heterocyclylalkyl, halogen,        cyano, nitro, —C(O)OR⁴, OR⁵, —NR^(a)R^(b), (NR^(a)R^(b))alkyl,        and (MeO)(HO)P(O)O—, and    -   wherein cycloalkyl and heterocyclyl may optionally be fused onto        an aromatic ring and may optionally be substituted with one or        more substituents independently selected from alkyl, hydroxyl,        halogen, aryl, —NR^(a)R^(b), oxo, and —C(O)OR⁴;    -   R³ is hydrogen, alkyl, or arylalkyl;    -   R⁴ is alkyl or arylalkyl;    -   R⁵ is hydrogen, alkyl, or arylalkyl;    -   R^(a) and R^(b) are independently selected from hydrogen, alkyl,        cycloalkyl, arylalkyl, heteroaryl, —C(O)R⁶, —C(O)OR⁷,        —C(O)NR^(c)R^(d), and (NR^(c)R^(d))alkyl, or alternatively,        R^(a) and R^(b), together with the nitrogen atom to which they        are attached, form a five- or six-membered ring or bridged        bicyclic ring structure, wherein said five- or six-membered ring        or bridged bicyclic ring structure optionally may contain one or        two additional heteroatoms independently selected from nitrogen,        oxygen, and sulfur and may contain one, two, or three        substituents independently selected from C₁ to C₆ alkyl, C₁ to        C₄ haloalkyl, aryl, hydroxyl, C₁ to C₆ alkoxy, C₁ to C₄        haloalkoxy, and halogen;    -   R⁶ is alkyl;    -   R⁷ is alkyl, arylalkyl, or haloalkyl; and    -   R^(c) and R^(d) are independently selected from hydrogen, alkyl,        arylalkyl, and cycloalkyl.

In a sixteenth embodiment of the first aspect, the present disclosureprovides a compound, or a pharmaceutically acceptable salt or solvatethereof, selected from the group consisting of:

-   (1R)-2-((2S)-2-(4-(4-(4-(2-((2S)-1-((2R)-2-(dimethylamino)-2-phenylacetyl)-2-pyrrolidinyl)-1H-imidazol-5-yl)phenoxy)phenyl)-1H-imidazol-2-yl)-1-pyrrolidinyl)-N,N-dimethyl-2-oxo-1-phenylethanamine;-   (1R)-2-((2S)-2-(4-(4-(4-(2-((2S)-1-((2R)-2-hydroxy-2-phenylacetyl)-2-pyrrolidinyl)-1H-imidazol-5-yl)phenoxy)phenyl)-1H-imidazol-2-yl)-1-pyrrolidinyl)-2-oxo-1-phenylethanol;-   dimethyl(oxybis(4,1-phenylene-1H-imidazole-4,2-diyl(2S)-2,1-pyrrolidinediyl((1R)-2-oxo-1-phenyl-2,1-ethanediyl)))biscarbamate;-   (1R)-2-((2S)-2-(4-(3-(4-(2-((2S)-1-((2R)-2-(dimethylamino)-2-phenylacetyl)-2-pyrrolidinyl)-1H-imidazol-4-yl)phenoxy)phenyl)-1H-imidazol-2-yl)-1-pyrrolidinyl)-N,N-dimethyl-2-oxo-1-phenylethanamine;-   (1R)-2-((2S)-2-(4-(3-(4-(2-((2S)-1-((2R)-2-hydroxy-2-phenylacetyl)-2-pyrrolidinyl)-1H-imidazol-4-yl)phenoxy)phenyl)-1H-imidazol-2-yl)-1-pyrrolidinyl)-2-oxo-1-phenylethanol;-   methyl((1R)-2-((2S)-2-(4-(3-(4-(2-((2S)-1-((2R)-2-((methoxycarbonyl)amino)-2-phenylacetyl)-2-pyrrolidinyl)-1H-imidazol-4-yl)phenoxy)phenyl)-1H-imidazol-2-yl)-1-pyrrolidinyl)-2-oxo-1-phenylethyl)carbamate;-   (1R)-2-((2S)-2-(4-(4-((4-(2-((2S)-1-((2R)-2-(dimethylamino)-2-phenylacetyl)-2-pyrrolidinyl)-1H-imidazol-4-yl)benzyl)oxy)phenyl)-1H-imidazol-2-yl)-1-pyrrolidinyl)-N,N-dimethyl-2-oxo-1-phenylethanamine;-   methyl((1R)-2-((2S)-2-(4-(4-((4-(2-((2S)-1-((2R)-2-((methoxycarbonyl)amino)-2-phenylacetyl)-2-pyrrolidinyl)-1H-imidazol-4-yl)benzyl)oxy)phenyl)-1H-imidazol-2-yl)-1-pyrrolidinyl)-2-oxo-1-phenylethyl)carbamate;-   (1R)-2-((2S)-2-(4-(4-(2-(4-(2-((2S)-1-((2R)-2-(dimethylamino)-2-phenylacetyl)-2-pyrrolidinyl)-1H-imidazol-5-yl)phenyl)ethyl)phenyl)-1H-imidazol-2-yl)-1-pyrrolidinyl)-N,N-dimethyl-2-oxo-1-phenylethanamine;-   (1R,1′R)-2,2′-(1,2-ethanediylbis(4,1-phenylene-1H-imidazole-4,2-diyl(2S)-2,1-pyrrolidinediyl))bis(2-oxo-1-phenylethanol);-   dimethyl    (1,2-ethanediylbis(4,1-phenylene-1H-imidazole-4,2-diyl(2S)-2,1-pyrrolidinediyl((1R)-2-oxo-1-phenyl-2,1-ethanediyl)))biscarbamate;-   N′,N′″-(1,2-ethanediylbis(4,1-phenylene-1H-imidazole-4,2-diyl(2S)-2,1-pyrrolidinediyl((1R)-2-oxo-1-phenyl-2,1-ethanediyl)))bis(1-ethylurea);-   1-cyclopentyl-3-((1R)-2-((2S)-2-(4-(4-(2-(4-(2-((2S)-1-((2R)-2-((cyclopentylcarbamoyl)amino)-2-phenylacetyl)-2-pyrrolidinyl)-1H-imidazol-5-yl)phenyl)ethyl)phenyl)-1H-imidazol-2-yl)-1-pyrrolidinyl)-2-oxo-1-phenylethyl)urea;-   (1R)-2-((2S)-2-(4-(4-(((4-(2-((2S)-1-((2R)-2-(dimethylamino)-2-phenylacetyl)-2-pyrrolidinyl)-1H-imidazol-5-yl)benzyl)oxy)methyl)phenyl)-1H-imidazol-2-yl)-1-pyrrolidinyl)-N,N-dimethyl-2-oxo-1-phenylethanamine;-   (1R)-2-((2S)-2-(4-(4-(((4-(2-((2S)-1-((2R)-2-hydroxy-2-phenylacetyl)-2-pyrrolidinyl)-1H-imidazol-5-yl)benzyl)oxy)methyl)phenyl)-1H-imidazol-2-yl)-1-pyrrolidinyl)-2-oxo-1-phenylethanol;-   dimethyl(oxybis(methylene-4,1-phenylene-1H-imidazole-4,2-diyl(2S)-2,1-pyrrolidinediyl((1R)-2-oxo-1-phenyl-2,1-ethanediyl)))biscarbamate;-   1-methyl-3-((1R)-2-((2S)-2-(4-(4-(((4-(2-((2S)-1-((2R)-2-((methylcarbamoyl)amino)-2-phenylacetyl)-2-pyrrolidinyl)-1H-imidazol-5-yl)benzyl)oxy)methyl)phenyl)-1H-imidazol-2-yl)-1-pyrrolidinyl)-2-oxo-1-phenylethyl)urea;-   1-ethyl-3-((1R)-2-((2S)-2-(4-(4-(((4-(2-((2S)-1-((2R)-2-((ethylcarbamoyl)amino)-2-phenylacetyl)-2-pyrrolidinyl)-1H-imidazol-5-yl)benzyl)oxy)methyl)phenyl)-1H-imidazol-2-yl)-1-pyrrolidinyl)-2-oxo-1-phenylethyl)urea;-   1-cyclopentyl-3-((1R)-2-((2S)-2-(4-(4-(((4-(2-((2S)-1-((2R)-2-((cyclopentylcarbamoyl)amino)-2-phenylacetyl)-2-pyrrolidinyl)-1H-imidazol-5-yl)benzyl)oxy)methyl)phenyl)-1H-imidazol-2-yl)-1-pyrrolidinyl)-2-oxo-1-phenylethyl)urea;-   (1R)-2-((2S)-2-(4-(3-(((4-(2-((2S)-1-((2R)-2-(dimethylamino)-2-phenylacetyl)-2-pyrrolidinyl)-1H-imidazol-4-yl)benzyl)oxy)methyl)phenyl)-1H-imidazol-2-yl)-1-pyrrolidinyl)-N,N-dimethyl-2-oxo-1-phenylethanamine;-   (1R)-2-((2S)-2-(4-(3-(((4-(2-((2S)-1-((2R)-2-hydroxy-2-phenylacetyl)-2-pyrrolidinyl)-1H-imidazol-4-yl)benzyl)oxy)methyl)phenyl)-1H-imidazol-2-yl)-1-pyrrolidinyl)-2-oxo-1-phenylethanol;-   (methyl((1R)-2-((2S)-2-(4-(3-(((4-(2-((2S)-1-((2R)-2-((methoxycarbonyl)amino)-2-phenylacetyl)-2-pyrrolidinyl)-1H-imidazol-4-yl)benzyl)oxy)methyl)phenyl)-1H-imidazol-2-yl)-1-pyrrolidinyl)-2-oxo-1-phenylethyl)carbamate;-   1-methyl-3-((1R)-2-((2S)-2-(4-(3-(((4-(2-((2S)-1-((2R)-2-((methylcarbamoyl)amino)-2-phenylacetyl)-2-pyrrolidinyl)-1H-imidazol-4-yl)benzyl)oxy)methyl)phenyl)-1H-imidazol-2-yl)-1-pyrrolidinyl)-2-oxo-1-phenylethyl)urea;-   1-ethyl-3-((1R)-2-((2S)-2-(4-(3-(((4-(2-((2S)-1-((2R)-2-((ethylcarbamoyl)amino)-2-phenylacetyl)-2-pyrrolidinyl)-1H-imidazol-4-yl)benzyl)oxy)methyl)phenyl)-1H-imidazol-2-yl)-1-pyrrolidinyl)-2-oxo-1-phenylethyl)urea;-   1-cyclopentyl-3-((1R)-2-((2S)-2-(4-(3-(((4-(2-((2S)-1-((2R)-2-((cyclopentylcarbamoyl)amino)-2-phenylacetyl)-2-pyrrolidinyl)-1H-imidazol-4-yl)benzyl)oxy)methyl)phenyl)-1H-imidazol-2-yl)-1-pyrrolidinyl)-2-oxo-1-phenylethyl)urea;-   dimethyl    (1,1′:4′,1″-terphenyl-4,4″-diylbis(1H-imidazole-4,2-diyl(2S)-2,1-pyrrolidinediyl((1R)-2-oxo-1-phenyl-2,1-ethanediyl)))biscarbamate;-   (1R)-2-((2S)-2-(4-(4″-(2-((2S)-1-((2R)-2-(dimethylamino)-2-phenylacetyl)-2-pyrrolidinyl)-1H-imidazol-5-yl)-1,1′:4′,1″-terphenyl-4-yl)-1H-imidazol-2-yl)-1-pyrrolidinyl)-N,N-dimethyl-2-oxo-1-phenylethanamine;-   methyl((1S)-1-(((2S)-2-(4-(4-(4-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amino)-3-methylbutanoyl)pyrrolidin-2-yl)-1H-imidazol-4-yl)phenyl)cyclohexyl)-1H-imidazol-2-yl)pyrrolidin-1-yl)carbonyl)-2-methylpropyl)carbamate;-   methyl((1S)-1-(1R,3S,5R)-3-(4-chloro-5-(4-(4-(4-chloro-2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-5-yl)bicyclo[2.2.2]oct-1-yl)phenyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)carbonyl)-2-methylpropyl)carbamate;    and    -   corresponding stereoisomers thereof.

In a second aspect the present disclosure provides a compositioncomprising a compound of Formula (I), or a pharmaceutically acceptablesalt thereof, and a pharmaceutically acceptable carrier, wherein:

-   -   s is 0 or 1;    -   L is -L¹-L²-, wherein L¹ and L² are independently selected from:

-   -   Y and Y′ are independently oxygen (O) or NH;    -   R¹ is hydrogen or —C(O)R^(x);    -   R² is hydrogen or —C(O)R^(y);    -   R^(x) and R^(y) are independently selected from cycloalkyl,        heteroaryl, heterocyclyl, alkoxy, and alkyl substituted with one        or more substituents independently selected from aryl, alkenyl,        cycloalkyl, heterocyclyl, heteroaryl, —OR³,    -   —C(O)OR⁴, —NR^(a)R^(b), and —C(O)NR^(c)R^(d),    -   wherein aryl and heteroaryl may optionally be substituted with        one or more substituents independently selected from alkyl,        haloalkyl, arylalkyl, heterocyclyl, heterocyclylalkyl, halogen,        cyano, nitro, —C(O)OR⁴, OR⁵, —NR^(a)R^(b), (NR^(a)R^(b))alkyl,        and (MeO)(HO)P(O)O—, and    -   wherein cycloalkyl and heterocyclyl may optionally be fused onto        an aromatic ring and may optionally be substituted with one or        more substituents independently selected from alkyl, hydroxyl,        halogen, aryl, —NR^(a)R^(b), oxo, and —C(O)OR⁴;    -   R³ is hydrogen, alkyl, or arylalkyl;    -   R⁴ is alkyl or arylalkyl;    -   R⁵ is hydrogen, alkyl, or arylalkyl;    -   R^(a) and R^(b) are independently selected from hydrogen, alkyl,        cycloalkyl, arylalkyl, heteroaryl, —C(O)R⁶, —C(O)OR⁷,        —C(O)NR^(c)R^(d), and (NR^(c)R^(d))alkyl, or alternatively,        R^(a) and R^(b), together with the nitrogen atom to which they        are attached, form a five- or six-membered ring or bridged        bicyclic ring structure, wherein said five- or six-membered ring        or bridged bicyclic ring structure optionally may contain one or        two additional heteroatoms independently selected from nitrogen,        oxygen, and sulfur and may contain one, two, or three        substituents independently selected from C₁ to C₆ alkyl, C₁ to        C₄ haloalkyl, aryl, hydroxyl, C₁ to C₆ alkoxy, C₁ to C₄        haloalkoxy, and halogen;    -   R⁶ is alkyl;    -   R⁷ is alkyl, arylalkyl, or haloalkyl;    -   R¹⁰⁰ and R¹¹⁰ are independently selected from hydrogen and halo;        and    -   R^(c) and R^(d) are independently selected from hydrogen, alkyl,        arylalkyl, and cycloalkyl.

In a first embodiment of the second aspect the composition furthercomprises at least one additional compound having anti-HCV activity.

In a second embodiment of the second aspect at least one of theadditional compounds is an interferon or a ribavirin.

In a third embodiment of the second aspect the interferon is selectedfrom interferon alpha 2B, pegylated interferon alpha, consensusinterferon, interferon alpha 2A, and lymphoblastoid interferon tau.

In a fourth embodiment of the second aspect the present disclosureprovides a composition comprising a compound of Formula (I), or apharmaceutically acceptable salt thereof, a pharmaceutically acceptablecarrier, and at least one additional compound having anti-HCV activity,wherein at least one of the additional compounds is selected frominterleukin 2, interleukin 6, interleukin 12, a compound that enhancesthe development of a type 1 helper T cell response, interfering RNA,anti-sense RNA, Imiqimod, ribavirin, an inosine 5′-monophospatedehydrogenase inhibitor, amantadine, and rimantadine.

In a fifth embodiment of the second aspect the present disclosureprovides a composition comprising a compound of Formula (I), or apharmaceutically acceptable salt thereof, a pharmaceutically acceptablecarrier, and at least one additional compound having anti-HCV activity,wherein at least one of the additional compounds is effective to inhibitthe function of a target selected from HCV metalloprotease, HCV serineprotease, HCV polymerase, HCV helicase, HCV NS4B protein, HCV entry, HCVassembly, HCV egress, HCV NS5A protein, and IMPDH for the treatment ofan HCV infection.

In a third aspect the present disclosure provides a method of treatingan HCV infection in a patient, comprising administering to the patient atherapeutically effective amount of a compound of Formula (I), or apharmaceutically acceptable salt thereof, wherein:

-   -   s is 0 or 1;    -   L is -L¹-L²-, wherein L¹ and L² are independently selected from:

-   -   Y and Y′ are independently oxygen (O) or NH;    -   R¹ is hydrogen or —C(O)R^(x);    -   R² is hydrogen or —C(O)R^(y);    -   R^(x) and R^(y) are independently selected from cycloalkyl,        heteroaryl, heterocyclyl, alkoxy, and alkyl substituted with one        or more substituents independently selected from aryl, alkenyl,        cycloalkyl, heterocyclyl, heteroaryl, —OR³,    -   —C(O)OR⁴, —NR^(a)R^(b), and —C(O)NR^(c)R^(d),    -   wherein aryl and heteroaryl may optionally be substituted with        one or more substituents independently selected from alkyl,        haloalkyl, arylalkyl, heterocyclyl, heterocyclylalkyl, halogen,        cyano, nitro, —C(O)OR⁴, OR⁵, —NR^(a)R^(b), (NR^(a)R^(b))alkyl,        and (MeO)(HO)P(O)O—, and    -   wherein cycloalkyl and heterocyclyl may optionally be fused onto        an aromatic ring and may optionally be substituted with one or        more substituents independently selected from alkyl, hydroxyl,        halogen, aryl, —NR^(a)R^(b), oxo, and —C(O)OR⁴;    -   R³ is hydrogen, alkyl, or arylalkyl;    -   R⁴ is alkyl or arylalkyl;    -   R⁵ is hydrogen, alkyl, or arylalkyl;    -   R^(a) and R^(b) are independently selected from hydrogen, alkyl,        cycloalkyl, arylalkyl, heteroaryl, —C(O)R⁶, —C(O)OR⁷,        —C(O)NR^(c)R^(d), and (NR^(c)R^(d))alkyl, or alternatively,        R^(a) and R^(b), together with the nitrogen atom to which they        are attached, form a five- or six-membered ring or bridged        bicyclic ring structure, wherein said five- or six-membered ring        or bridged bicyclic ring structure optionally may contain one or        two additional heteroatoms independently selected from nitrogen,        oxygen, and sulfur and may contain one, two, or three        substituents independently selected from C₁ to C₆ alkyl, C₁ to        C₄ haloalkyl, aryl, hydroxyl, C₁ to C₆ alkoxy, C₁ to C₄        haloalkoxy, and halogen;    -   R⁶ is alkyl;    -   R⁷ is alkyl, arylalkyl, or haloalkyl; and    -   R^(c) and R^(d) are independently selected from hydrogen, alkyl,        arylalkyl, and cycloalkyl.

In a first embodiment of the third aspect the method further comprisesadministering at least one additional compound having anti-HCV activityprior to, after or simultaneously with the compound of Formula (I), or apharmaceutically acceptable salt thereof.

In a second embodiment of the third aspect at least one of theadditional compounds is an interferon or a ribavirin.

In a third embodiment of the third aspect the interferon is selectedfrom interferon alpha 2B, pegylated interferon alpha, consensusinterferon, interferon alpha 2A, and lymphoblastoid interferon tau.

In a fourth embodiment of the third aspect the present disclosureprovides a method of treating an HCV infection in a patient, comprisingadministering to the patient a therapeutically effective amount of acompound of Formula (I), or a pharmaceutically acceptable salt thereof,and at least one additional compound having anti-HCV activity prior to,after or simultaneously with the compound of Formula (I), or apharmaceutically acceptable salt thereof, wherein at least one of theadditional compounds is selected from interleukin 2, interleukin 6,interleukin 12, a compound that enhances the development of a type 1helper T cell response, interfering RNA, anti-sense RNA, Imiqimod,ribavirin, an inosine 5′-monophospate dehydrogenase inhibitor,amantadine, and rimantadine.

In a fifth embodiment of the third aspect the present disclosureprovides a method of treating an HCV infection in a patient, comprisingadministering to the patient a therapeutically effective amount of acompound of Formula (I), or a pharmaceutically acceptable salt thereof,and at least one additional compound having anti-HCV activity prior to,after or simultaneously with the compound of Formula (I), or apharmaceutically acceptable salt thereof, wherein at least one of theadditional compounds is effective to inhibit the function of a targetselected from HCV metalloprotease, HCV serine protease, HCV polymerase,HCV helicase, HCV NS4B protein, HCV entry, HCV assembly, HCV egress, HCVNS5A protein, and IMPDH for the treatment of an HCV infection.

Other aspects of the present disclosure may include suitablecombinations of embodiments disclosed herein.

Yet other aspects and embodiments may be found in the descriptionprovided herein.

The description of the present disclosure herein should be construed incongruity with the laws and principals of chemical bonding. In someinstances it may be necessary to remove a hydrogen atom in order toaccommodate a substituent at any given location.

Certain features of the structure of Formula (I) are further illustratedbelow:

In Formula (I), as depicted above, the “pyrrolidine moiety” on the leftside of the “linker” is independent from the “pyrrolidine moiety” on theright side in respect to, e.g., (1) tautomer form of imidazole ring whenY or Y′ is NH, (2) absolute configuration of the stereogenic centers onthe pyrrolidine ring, and (3) substituents on the pyrrolidine nitrogen,i.e., R¹ and R² are independent from each other, although in somecircumstances they are preferably the same.

As for connection between the linker “L” and the pyrrolidine moieties,Formula (I) encompasses all the following possible combinations:

wherein Y and Y′ are independently oxygen (O) or NH.

In a pyrrolidine ring, the stereogenic carbon center to which afive-membered heterocycle is attached can take either (R)- or(S)-configuration as depicted below:

When a cyclopropyl ring is fused onto a pyrrolidine ring, i.e., when sis 1, the CH₂ group of the fused cyclopropyl ring can take either α- orβ-position relative to the pyrrolidine ring, as depicted below:

In Formula (I), when either Y or Y′ is NH, the linkage between thelinker “L” and the resultant imidazole ring can take place in either theC-4 or the C-5 position (see below) of the imidazole ring. As a personof ordinary skill in the art would understand, due to tautomerization ofthe imidazole ring, a bonding of the linker “L” to the C-4 position maybe equivalent to a bonding of the linker to the C-5 position, as shownin the following equation:

Thus, this disclosure is intended to cover both possible linkages evenwhen a structure depicts only one of them.

In this disclosure, a floating bond (e.g.,

or a floating substituent (e.g., —R¹³) on a structure indicates that thebond or substituent can attach to any available position of thestructure by removal of a hydrogen from the available position. Itshould be understood that in a bicyclic or polycyclic ring structure,unless specifically defined otherwise, the position of a floating bondor a floating substituent does not limit the position of such bond orsubstituent to a specific ring. Thus, the following two substituentgroups should be construed to be equivalent:

It should be understood that the compounds encompassed by the presentdisclosure are those that are suitably stable for use as pharmaceuticalagent.

It is intended that the definition of any substituent or variable at aparticular location in a molecule be independent of its definitionselsewhere in that molecule. For example, for substituent (R¹⁰)_(n), whenn is 2, each of the two R¹⁰ groups may be the same or different.

All patents, patent applications, and literature references cited in thespecification are herein incorporated by reference in their entirety. Inthe case of inconsistencies, the present disclosure, includingdefinitions, will prevail.

DEFINITIONS

Definitions have been provided above for each of the groups defined. Inaddition, the following definitions shall be used.

As used herein, the singular forms “a”, “an”, and “the” include pluralreference unless the context clearly dictates otherwise.

Unless stated otherwise, all aryl, cycloalkyl, heteroaryl, andheterocyclyl groups of the present disclosure may be substituted asdescribed in each of their respective definitions. For example, the arylpart of an arylalkyl group may be substituted as described in thedefinition of the term “aryl.”

The term “acetyl,” as used herein, refers to —C(O)CH₃.

The term “alkenyl,” as used herein, refers to a monovalent, straight orbranched hydrocarbon chain having one or more double bonds therein. Thedouble bond of an alkenyl group can be unconjugated or conjugated toanother unsaturated group. Suitable alkenyl groups include, but are notlimited to, C₂ to C₁₀ alkenyl groups, such as vinyl, allyl, butenyl,pentenyl, hexenyl, butadienyl, pentadienyl, hexadienyl, 2-ethylhexenyl,2-propyl-2-butenyl, 4-(2-methyl-3-butene)-pentenyl. An alkenyl group canbe unsubstituted or substituted with one or two suitable substituents.

The term “alkoxy,” as used herein, refers to an alkyl group attached tothe parent molecular moiety through an oxygen atom. Representativeexamples of alkoxy group include, but are not limited to, methoxy(CH₃O—), ethoxy (CH₃CH₂O—), and t-butoxy ((CH₃)₃CO—).

The term “alkyl,” as used herein, refers to a group derived from astraight or branched chain saturated hydrocarbon by removal of ahydrogen from one of the saturated carbons. The alkyl group preferablycontains from one to ten carbon atoms. Representative examples of alkylgroup include, but are not limited to, methyl, ethyl, isopropyl, andtert-butyl.

The term “alkylcarbonyl,” as used herein, refers to an alkyl groupattached to the parent molecular moiety through a carbonyl group.Representative examples of alkylcarbonyl group include, but are notlimited to, acetyl (—C(O)CH₃), propanoyl (—C(O)CH₂CH₃), n-butyryl(—C(O)CH₂CH₂CH₃), and 2,2-dimethylpropanoyl or pivaloyl (—C(O)C(CH₃)₃).

The term “allyl,” as used herein, refers to the —CH₂CH═CH₂ group.

The term “aryl,” as used herein, refers to a group derived from anaromatic carbocycle by removal of a hydrogen atom from an aromatic ring.The aryl group can be monocyclic, bicyclic or polycyclic, wherein inbicyclic or polycyclic aryl group, the aromatic carbocycle can be fusedonto another four- to six-membered aromatic or non-aromatic carbocycle.Representative examples of aryl groups include, but are not limited to,phenyl, indanyl, indenyl, naphthyl, and 1,2,3,4-tetrahydronaphth-5-yl.

The term “arylalkyl,” as used herein, refers to an alkyl groupsubstituted with one, two, or three aryl groups, wherein aryl part ofthe arylalkyl group may optionally be substituted by one to fivesubstituents independently selected from C₁ to C₆ alkyl, C₁ to C₄haloalkyl, C₁ to C₆ alkoxy, halogen, cyano, and nitro groups.Represented examples of arylalkyl include, but are not limited to,benzyl, 2-phenyl-1-ethyl(PhCH₂CH₂—), (naphth-1-yl)methyl, and(naphth-2-yl)methyl.

The term “benzyl,” as used herein, refers to a methyl group on which oneof the hydrogen atoms is replaced by a phenyl group, wherein said phenylgroup may optionally be substituted by one to five substituentsindependently selected from methyl, trifluoromethyl (—CF₃), methoxy(—OCH₃), halogen, and nitro (—NO₂). Representative examples of benzylgroup include, but are not limited to, PhCH₂—, 4-MeO—C₆H₄CH₂—, and2,4,6-tri-methyl-C₆H₄CH₂—.

The terms “Cap” and “cap,” as used herein, refer to the group which isplaced on the nitrogen atom of the pyrrolidine ring in the compounds offormula (I). It should be understood that “Cap” or “cap” can also referto the reagent which is a precursor to the final “cap” in compounds offormula (I) and is used as one of the starting materials in the reactionto append a group on the pyrrolidine nitrogen that results in the finalproduct, a compound which contains the functionalized pyrrolidine thatwill be present in the compound of formula (I).

The term “carbonyl,” as used herein, refers to —C(O)—.

The term “carboxyl,” as used herein, refers to —CO₂H.

The term “cyano,” as used herein, refers to —CN.

The term “cycloalkyl,” as used herein, refers to a group derived from asaturated carbocycle, having preferably three to eight carbon atoms, byremoval of a hydrogen atom from the saturated carbocycle, wherein thesaturated carbocycle can optionally be fused onto one or two otheraromatic or nonaromatic carbocycles. Representative examples ofcycloalkyl groups include, but are not limited to, cyclopropyl,cyclopentyl, cyclohexyl, and 1,2,3,4-tetrahydronaphth-1-yl.

The term “formyl,” as used herein, refers to —CHO.

The terms “halo” and “halogen,” as used herein, refer to F, Cl, Br, orI.

The term “haloalkoxy,” as used herein, refers to a haloalkyl groupattached to the parent molecular moiety through an oxygen atom.

The term “haloalkyl,” as used herein, refers to an alkyl groupsubstituted by at least one halogen atom. The haloalkyl group can be analkyl group of which all hydrogen atoms are substituted by halogens.Representative examples of haloalkyl include, but are not limited to,trifluoromethyl (CF₃—), 1-chloroethyl (ClCH₂CH₂—), and2,2,2-trifluoroethyl (CF₃CH₂—).

The term “heteroaryl,” as used herein, refers to group derived from amonocyclic, bicyclic, or polycyclic compound comprising at least onearomatic ring comprising one or more heteroatoms, preferably, one tothree heteroatoms, independently selected from nitrogen, oxygen, andsulfur, by removal of a hydrogen atom from an aromatic ring thereof. Asis well known to those skilled in the art, heteroaryl rings have lessaromatic character than their all-carbon counterparts. Thus, for thepurposes of the invention, a heteroaryl group need only have some degreeof aromatic character. Illustrative examples of heteroaryl groupsinclude, but are not limited to, pyridyl, pyridazinyl, pyrimidyl,pyrazyl, triazinyl, pyrrolyl, pyrazolyl, imidazolyl, (1,2,3,)- and(1,2,4)-triazolyl, pyrazinyl, pyrimidinyl, tetrazolyl, furyl, thienyl,isoxazolyl, thiazolyl, isoxazolyl, oxazolyl, indolyl, quinolinyl,isoquinolinyl, benzisoxazolyl, benzothiazolyl, benzothienyl, andpyrrolopyridinyl.

The term “heteroarylalkyl,” as used herein, refers to an alkyl groupsubstituted with one, two, or three heteroaryl groups.

The term “heterobicyclyl,” as used herein, refers to a ring structurecomprising two fused or bridged rings that include carbon and one ormore heteroatoms independently selected from nitrogen, oxygen, andsulfur as the ring atom(s). The heterobicyclic ring structure is asubset of heterocyclic ring and can be saturated or unsaturated.Examples of heterobicyclic ring structures include tropane,quinuclidine, and 7-azabicyclo[2.2.1]heptane.

The term “heterocyclyl,” as used herein, refers to a group derived froma monocyclic, bicyclic, or polycyclic compound comprising at least onenonaromatic ring comprising one or more heteroatoms, preferably, one tothree heteroatoms, independently selected from nitrogen, oxygen, andsulfur, by removal of a hydrogen atom from the nonaromatic ring. Theheterocyclyl group encompasses the heterobicyclyl group. Theheterocyclyl groups of the present disclosure can be attached to theparent molecular moiety through a carbon atom or a nitrogen atom in thegroup. Examples of heterocyclyl groups include, but are not limited to,morpholinyl, oxazolidinyl, piperazinyl, piperidinyl, pyrrolidinyl,tetrahydrofuryl, thiomorpholinyl, and indolinyl.

The term “heterocyclylalkyl,” as used herein, refers to an alkyl groupsubstituted with one, two, or three heterocyclyl groups.

The terms “hydroxy” or “hydroxyl,” as used herein, refer to —OH.

The term “nitro,” as used herein, refers to —NO₂.

The term “—NR^(a)R^(b),” as used herein, refers to two groups, R^(a) andR^(b), which are attached to the parent molecular moiety through anitrogen atom, or alternatively R^(a) and R^(b), together with thenitrogen atom to which they are attached, form a 5- or 6-membered ringor a fused- or bridged-bicyclic ring structure optionally containingone, two, or three additional heteroatom independently selected fromnitrogen, oxygen, and sulfur. The term “—NR^(c)R^(d)” is definedsimilarly.

The term “(NR^(a)R^(b))alkyl,” as used herein, refers to an alkyl groupsubstituted with one, two, or three —NR^(a)R^(b) groups. The term“(NR^(c)R^(d))alkyl” is defined similarly.

The term “oxo,” as used herein, refers to ═O.

The term “sulfonyl,” as used herein, refers to —SO₂—.

In the compounds of the present disclosure, it should be understood thatwhen is 0, the compound of formula (Z) shown below is the result:

Asymmetric centers exist in the compounds of the present disclosure.These centers are designated by the symbols “R” or “S”, depending on theconfiguration of substituents around the chiral carbon atom. It shouldbe understood that the disclosure encompasses all stereochemicalisomeric forms, or mixtures thereof, which possess the ability toinhibit NS5A. Individual stereoisomers of compounds can be preparedsynthetically from commercially available starting materials whichcontain chiral centers or by preparation of mixtures of enantiomericproducts followed by separation such as conversion to a mixture ofdiastereomers followed by separation or recrystallization,chromatographic techniques, or direct separation of enantiomers onchiral chromatographic columns. Starting compounds of particularstereochemistry are either commercially available or can be made andresolved by techniques known in the art.

Certain compounds of the present disclosure may also exist in differentstable conformational forms which may be separable. Torsional asymmetrydue to restricted rotation about an asymmetric single bond, for examplebecause of steric hindrance or ring strain, may permit separation ofdifferent conformers. The present disclosure includes eachconformational isomer of these compounds and mixtures thereof.

The term “compounds of the present disclosure”, and equivalentexpressions, are meant to embrace compounds of Formula (I), andpharmaceutically acceptable enantiomers, diastereomers, and saltsthereof. Similarly, references to intermediates are meant to embracetheir salts where the context so permits.

The present disclosure is intended to include all isotopes of atomsoccurring in the present compounds. Isotopes include those atoms havingthe same atomic number but different mass numbers. By way of generalexample and without limitation, isotopes of hydrogen include deuteriumand tritium. Isotopes of carbon include ¹³C and ¹⁴C.Isotopically-labeled compounds of the invention can generally beprepared by conventional techniques known to those skilled in the art orby processes analogous to those described herein, using an appropriateisotopically-labeled reagent in place of the non-labeled reagentotherwise employed. Such compounds may have a variety of potential uses,for example as standards and reagents in determining biologicalactivity. In the case of stable isotopes, such compounds may have thepotential to favorably modify biological, pharmacological, orpharmacokinetic properties.

The compounds of the present disclosure can exist as pharmaceuticallyacceptable salts. The term “pharmaceutically acceptable salt,” as usedherein, represents salts or zwitterionic forms of the compounds of thepresent disclosure which are water or oil-soluble or dispersible, whichare, within the scope of sound medical judgment, suitable for use incontact with the tissues of patients without excessive toxicity,irritation, allergic response, or other problem or complicationcommensurate with a reasonable benefit/risk ratio, and are effective fortheir intended use The salts can be prepared during the final isolationand purification of the compounds or separately by reacting a suitablenitrogen atom with a suitable acid. Representative acid addition saltsinclude acetate, adipate, alginate, citrate, aspartate, benzoate,benzenesulfonate, bisulfate, butyrate, camphorate, camphorsulfonate;digluconate, glycerophosphate, hemisulfate, heptanoate, hexanoate,formate, fumarate, hydrochloride, hydrobromide, hydroiodide,2-hydroxyethanesulfonate, lactate, maleate, mesitylenesulfonate,methanesulfonate, naphthylenesulfonate, nicotinate,2-naphthalenesulfonate, oxalate, palmoate, pectinate, persulfate,3-phenylproprionate, picrate, pivalate, propionate, succinate, tartrate,trichloroacetate, trifluoroacetate, phosphate, glutamate, bicarbonate,para-toluenesulfonate, and undecanoate. Examples of acids which can beemployed to form pharmaceutically acceptable addition salts includeinorganic acids such as hydrochloric, hydrobromic, sulfuric, andphosphoric, and organic acids such as oxalic, maleic, succinic, andcitric.

Basic addition salts can be prepared during the final isolation andpurification of the compounds by reacting a carboxy group with asuitable base such as the hydroxide, carbonate, or bicarbonate of ametal cation or with ammonia or an organic primary, secondary, ortertiary amine. The cations of pharmaceutically acceptable salts includelithium, sodium, potassium, calcium, magnesium, and aluminum, as well asnontoxic quaternary amine cations such as ammonium, tetramethylammonium,tetraethylammonium, methylamine, dimethylamine, trimethylamine,triethylamine, diethylamine, ethylamine, tributylamine, pyridine,N,N-dimethylaniline, N-methylpiperidine, N-methylmorpholine,dicyclohexylamine, procaine, dibenzylamine, N,N-dibenzylphenethylamine,and N,N′-dibenzylethylenediamine. Other representative organic aminesuseful for the formation of base addition salts include ethylenediamine,ethanolamine, diethanolamine, piperidine, and piperazine.

When it is possible that, for use in therapy, therapeutically effectiveamounts of a compound of Formula (I), as well as pharmaceuticallyacceptable salts thereof, may be administered as the raw chemical, it ispossible to present the active ingredient as a pharmaceuticalcomposition. Accordingly, the disclosure further provides pharmaceuticalcompositions, which include therapeutically effective amounts ofcompounds of Formula (I) or pharmaceutically acceptable salts thereof,and one or more pharmaceutically acceptable carriers, diluents, orexcipients. The term “therapeutically effective amount,” as used herein,refers to the total amount of each active component that is sufficientto show a meaningful patient benefit, e.g., a sustained reduction inviral load. When applied to an individual active ingredient,administered alone, the term refers to that ingredient alone. Whenapplied to a combination, the term refers to combined amounts of theactive ingredients that result in the therapeutic effect, whetheradministered in combination, serially, or simultaneously. The compoundsof Formula (I) and pharmaceutically acceptable salts thereof, are asdescribed above. The carrier(s), diluent(s), or excipient(s) must beacceptable in the sense of being compatible with the other ingredientsof the formulation and not deleterious to the recipient thereof. Inaccordance with another aspect of the present disclosure there is alsoprovided a process for the preparation of a pharmaceutical formulationincluding admixing a compound of Formula (I), or a pharmaceuticallyacceptable salt thereof, with one or more pharmaceutically acceptablecarriers, diluents, or excipients. The term “pharmaceuticallyacceptable,” as used herein, refers to those compounds, materials,compositions, and/or dosage forms which are, within the scope of soundmedical judgment, suitable for use in contact with the tissues ofpatients without excessive toxicity, irritation, allergic response, orother problem or complication commensurate with a reasonablebenefit/risk ratio, and are effective for their intended use.

Pharmaceutical formulations may be presented in unit dose formscontaining a predetermined amount of active ingredient per unit dose.Dosage levels of between about 0.01 and about 250 milligram per kilogram(“mg/kg”) body weight per day, preferably between about 0.05 and about100 mg/kg body weight per day of the compounds of the present disclosureare typical in a monotherapy for the prevention and treatment of HCVmediated disease. Typically, the pharmaceutical compositions of thisdisclosure will be administered from about 1 to about 5 times per day oralternatively, as a continuous infusion. Such administration can be usedas a chronic or acute therapy. The amount of active ingredient that maybe combined with the carrier materials to produce a single dosage formwill vary depending on the condition being treated, the severity of thecondition, the time of administration, the route of administration, therate of excretion of the compound employed, the duration of treatment,and the age, gender, weight, and condition of the patient. Preferredunit dosage formulations are those containing a daily dose or sub-dose,as herein above recited, or an appropriate fraction thereof, of anactive ingredient. Generally, treatment is initiated with small dosagessubstantially less than the optimum dose of the compound. Thereafter,the dosage is increased by small increments until the optimum effectunder the circumstances is reached. In general, the compound is mostdesirably administered at a concentration level that will generallyafford antivirally effective results without causing any harmful ordeleterious side effects.

When the compositions of this disclosure comprise a combination of acompound of the present disclosure and one or more additionaltherapeutic or prophylactic agent, both the compound and the additionalagent are usually present at dosage levels of between about 10 to 150%,and more preferably between about 10 and 80% of the dosage normallyadministered in a monotherapy regimen.

Pharmaceutical formulations may be adapted for administration by anyappropriate route, for example by the oral (including buccal orsublingual), rectal, nasal, topical (including buccal, sublingual, ortransdermal), vaginal, or parenteral (including subcutaneous,intracutaneous, intramuscular, intra-articular, intrasynovial,intrasternal, intrathecal, intralesional, intravenous, or intradermalinjections or infusions) route. Such formulations may be prepared by anymethod known in the art of pharmacy, for example by bringing intoassociation the active ingredient with the carrier(s) or excipient(s).Oral administration or administration by injection are preferred.

Pharmaceutical formulations adapted for oral administration may bepresented as discrete units such as capsules or tablets; powders orgranules; solutions or suspensions in aqueous or non-aqueous liquids;edible foams or whips; or oil-in-water liquid emulsions or water-in-oilemulsions.

For instance, for oral administration in the form of a tablet orcapsule, the active drug component can be combined with an oral,non-toxic pharmaceutically acceptable inert carrier such as ethanol,glycerol, water, and the like. Powders are prepared by comminuting thecompound to a suitable fine size and mixing with a similarly comminutedpharmaceutical carrier such as an edible carbohydrate, as, for example,starch or mannitol. Flavoring, preservative, dispersing, and coloringagent can also be present.

Capsules are made by preparing a powder mixture, as described above, andfilling formed gelatin sheaths. Glidants and lubricants such ascolloidal silica, talc, magnesium stearate, calcium stearate, or solidpolyethylene glycol can be added to the powder mixture before thefilling operation. A disintegrating or solubilizing agent such asagar-agar, calcium carbonate, or sodium carbonate can also be added toimprove the availability of the medicament when the capsule is ingested.

Moreover, when desired or necessary, suitable binders, lubricants,disintegrating agents, and coloring agents can also be incorporated intothe mixture. Suitable binders include starch, gelatin, natural sugarssuch as glucose or beta-lactose, corn sweeteners, natural and syntheticgums such as acacia, tragacanth or sodium alginate,carboxymethylcellulose, polyethylene glycol, and the like. Lubricantsused in these dosage forms include sodium oleate, sodium chloride, andthe like. Disintegrators include, without limitation, starch, methylcellulose, agar, betonite, xanthan gum, and the like. Tablets areformulated, for example, by preparing a powder mixture, granulating orslugging, adding a lubricant and disintegrant, and pressing intotablets. A powder mixture is prepared by mixing the compound, suitablecomminuted, with a diluent or base as described above, and optionally,with a binder such as carboxymethylcellulose, an aliginate, gelating, orpolyvinyl pyrrolidone, a solution retardant such as paraffin, aresorption accelerator such as a quaternary salt and/or and absorptionagent such as betonite, kaolin, or dicalcium phosphate. The powdermixture can be granulated by wetting with a binder such as syrup, starchpaste, acadia mucilage, or solutions of cellulosic or polymericmaterials and forcing through a screen. As an alternative togranulating, the powder mixture can be run through the tablet machineand the result is imperfectly formed slugs broken into granules. Thegranules can be lubricated to prevent sticking to the tablet formingdies by means of the addition of stearic acid, a stearate salt, talc, ormineral oil. The lubricated mixture is then compressed into tablets. Thecompounds of the present disclosure can also be combined with a freeflowing inert carrier and compressed into tablets directly without goingthrough the granulating or slugging steps. A clear or opaque protectivecoating consisting of a sealing coat of shellac, a coating of sugar orpolymeric material, and a polish coating of wax can be provided.Dyestuffs can be added to these coatings to distinguish different unitdosages.

Oral fluids such as solution, syrups, and elixirs can be prepared indosage unit form so that a given quantity contains a predeterminedamount of the compound. Syrups can be prepared by dissolving thecompound in a suitably flavored aqueous solution, while elixirs areprepared through the use of a non-toxic vehicle. Solubilizers andemulsifiers such as ethoxylated isostearyl alcohols and polyoxyethylenesorbitol ethers, preservatives, flavor additive such as peppermint oilor natural sweeteners, or saccharin or other artificial sweeteners, andthe like can also be added.

Where appropriate, dosage unit formulations for oral administration canbe microencapsulated. The formulation can also be prepared to prolong orsustain the release as for example by coating or embedding particulatematerial in polymers, wax, or the like.

The compounds of Formula (I), and pharmaceutically acceptable saltsthereof, can also be administered in the form of liposome deliverysystems, such as small unilamellar vesicles, large unilamellar vesicles,and multilamellar vesicles. Liposomes can be formed from a variety ofphospholipids, such as cholesterol, stearylamine, orphophatidylcholines.

The compounds of Formula (I) and pharmaceutically acceptable saltsthereof may also be delivered by the use of monoclonal antibodies asindividual carriers to which the compound molecules are coupled. Thecompounds may also be coupled with soluble polymers as targetable drugcarriers. Such polymers can include polyvinylpyrrolidone, pyrancopolymer, polyhydroxypropylmethacrylamidephenol,polyhydroxyethylaspartamidephenol, or polyethyleneoxidepolylysinesubstituted with palitoyl residues. Furthermore, the compounds may becoupled to a class of biodegradable polymers useful in achievingcontrolled release of a drug, for example, polylactic acid, polepsiloncaprolactone, polyhydroxy butyric acid, polyorthoesters, polyacetals,polydihydropyrans, polycyanoacrylates, and cross-linked or amphipathicblock copolymers of hydrogels.

Pharmaceutical formulations adapted for transdermal administration maybe presented as discrete patches intended to remain in intimate contactwith the epidermis of the recipient for a prolonged period of time. Forexample, the active ingredient may be delivered from the patch byiontophoresis as generally described in Pharm. Res., 3(6):318 (1986).

Pharmaceutical formulations adapted for topical administration may beformulated as ointments, creams, suspensions, lotions, powders,solutions, pastes, gels, sprays, aerosols, or oils.

For treatments of the eye or other external tissues, for example mouthand skin, the formulations are preferably applied as a topical ointmentor cream. When formulated in an ointment, the active ingredient may beemployed with either a paraffinic or a water-miscible ointment base.Alternatively, the active ingredient may be formulated in a cream withan oil-in-water cream base or a water-in oil base.

Pharmaceutical formulations adapted for topical administrations to theeye include eye drops wherein the active ingredient is dissolved orsuspended in a suitable carrier, especially an aqueous solvent.

Pharmaceutical formulations adapted for topical administration in themouth include lozenges, pastilles, and mouth washes.

Pharmaceutical formulations adapted for rectal administration may bepresented as suppositories or as enemas.

Pharmaceutical formulations adapted for nasal administration wherein thecarrier is a solid include a course powder having a particle size forexample in the range 20 to 500 microns which is administered in themanner in which snuff is taken, i.e., by rapid inhalation through thenasal passage from a container of the powder held close up to the nose.Suitable formulations wherein the carrier is a liquid, foradministration as a nasal spray or nasal drops, include aqueous or oilsolutions of the active ingredient.

Pharmaceutical formulations adapted for administration by inhalationinclude fine particle dusts or mists, which may be generated by means ofvarious types of metered, dose pressurized aerosols, nebulizers, orinsufflators.

Pharmaceutical formulations adapted for vaginal administration may bepresented as pessaries, tampons, creams, gels, pastes, foams, or sprayformulations.

Pharmaceutical formulations adapted for parenteral administrationinclude aqueous and non-aqueous sterile injection solutions which maycontain anti-oxidants, buffers, bacteriostats, and soutes which renderthe formulation isotonic with the blood of the intended recipient; andaqueous and non-aqueous sterile suspensions which may include suspendingagents and thickening agents. The formulations may be presented inunit-dose or multi-dose containers, for example sealed ampoules andvials, and may be stored in a freeze-dried (lyophilized) conditionrequiring only the addition of the sterile liquid carrier, for examplewater for injections, immediately prior to use. Extemporaneous injectionsolutions and suspensions may be prepared from sterile powders,granules, and tablets.

It should be understood that in addition to the ingredients particularlymentioned above, the formulations may include other agents conventionalin the art having regard to the type of formulation in question, forexample those suitable for oral administration may include flavoringagents.

The term “patient” includes both human and other mammals.

The term “treating” refers to: (i) preventing a disease, disorder orcondition from occurring in a patient that may be predisposed to thedisease, disorder, and/or condition but has not yet been diagnosed ashaving it; (ii) inhibiting the disease, disorder, or condition, i.e.,arresting its development; and (iii) relieving the disease, disorder, orcondition, i.e., causing regression of the disease, disorder, and/orcondition.

The compounds of the present disclosure can also be administered with acyclosporin, for example, cyclosporin A. Cyclosporin A has been shown tobe active against HCV in clinical trials (Hepatology, 38:1282 (2003);Biochem. Biophys. Res. Commun., 313:42 (2004); J. Gastroenterol., 38:567(2003)).

Table 1 below lists some illustrative examples of compounds that can beadministered with the compounds of this disclosure. The compounds of thedisclosure can be administered with other anti-HCV activity compounds incombination therapy, either jointly or separately, or by combining thecompounds into a composition.

TABLE 1 Physiological Type of Inhibitor or Brand Name Class TargetSource Company NIM811 Cyclophilin Novartis Inhibitor ZadaxinImmuno-modulator Sciclone Suvus Methylene blue Bioenvision Actilon TLR9agonist Coley (CPG10101) Batabulin (T67) Anticancer β-Tubulin inhibitorTularik Inc., South San Francisco, CA ISIS 14803 Antiviral AntisenseISIS Pharmaceuticals Inc, Carlsbad, CA/Elan Phamaceuticals Inc., NewYork, NY Summetrel Antiviral Antiviral Endo Pharmaceuticals HoldingsInc., Chadds Ford, PA GS-9132 (ACH-806) Antiviral HCV inhibitorAchillion/Gilead Pyrazolopyrimidine Antiviral HCV inhibitors Arrowcompounds and salts Therapeutics Ltd. From WO2005/047288 26 May 2005Levovirin Antiviral IMPDH inhibitor Ribapharm Inc., Costa Mesa, CAMerimepodib Antiviral IMPDH inhibitor Vertex Pharmaceuticals (VX-497)Inc., Cambridge, MA XTL-6865 (XTL- Antiviral Monoclonal XTL 002)antibody Biopharmaceuticals Ltd., Rehovot, Israel Telaprevir AntiviralNS3 serine protease Vertex Pharmaceuticals (VX-950, LY- inhibitor Inc.,Cambridge, 570310) MA/Eli Lilly and Co. Inc., Indianapolis, IN HCV-796Antiviral NS5B replicase Wyeth/ inhibitor Viropharma NM-283 AntiviralNS5B replicase Idenix/Novartis inhibitor GL-59728 Antiviral NS5Breplicase Gene Labs/ inhibitor Novartis GL-60667 Antiviral NS5Breplicase Gene Labs/ inhibitor Novartis 2′C MeA Antiviral NS5B replicaseGilead inhibitor PSI 6130 Antiviral NS5B replicase Roche inhibitor R1626Antiviral NS5B replicase Roche inhibitor 2′C Methyl Antiviral NS5Breplicase Merck adenosine inhibitor JTK-003 Antiviral RdRp inhibitorJapan Tobacco Inc., Tokyo, Japan Levovirin Antiviral Ribavirin ICNPharmaceuticals, Costa Mesa, CA Ribavirin Antiviral RibavirinSchering-Plough Corporation, Kenilworth, NJ Viramidine AntiviralRibavirin prodrug Ribapharm Inc., Costa Mesa, CA Heptazyme AntiviralRibozyme Ribozyme Pharmaceuticals Inc., Boulder, CO BILN-2061 AntiviralSerine protease Boehringer inhibitor Ingelheim Pharma KG, Ingelheim,Germany SCH 503034 Antiviral Serine protease Schering Plough inhibitorZadazim Immune modulator Immune modulator SciClone Pharmaceuticals Inc.,San Mateo, CA Ceplene Immunomodulator Immune modulator MaximPharmaceuticals Inc., San Diego, CA CellCept Immunosuppressant HCV IgGimmuno- F. Hoffmann-La suppressant Roche LTD, Basel, Switzerland CivacirImmunosuppressant HCV IgG immuno- Nabi suppressant BiopharmaceuticalsInc., Boca Raton, FL Albuferon - α Interferon Albumin IFN-α2b HumanGenome Sciences Inc., Rockville, MD Infergen A Interferon IFN InterMunealfacon-1 Pharmaceuticals Inc., Brisbane, CA Omega IFN Interferon IFN-ωIntarcia Therapeutics IFN-β and Interferon IFN-β and EMZ701 TransitionEMZ701 Therapeutics Inc., Ontario, Canada Rebif Interferon IFN-β1aSerono, Geneva, Switzerland Roferon A Interferon IFN-α2a F. Hoffmann-LaRoche LTD, Basel, Switzerland Intron A Interferon IFN-α2bSchering-Plough Corporation, Kenilworth, NJ Intron A and InterferonIFN-α2b/α1- RegeneRx Zadaxin thymosin Biopharma. Inc., Bethesda, MD/SciClone Pharmaceuticals Inc, San Mateo, CA Rebetron InterferonIFN-α2b/ribavirin Schering-Plough Corporation, Kenilworth, NJ ActimmuneInterferon INF-γ InterMune Inc., Brisbane, CA Interferon-β InterferonInterferon-β-1a Serono Multiferon Interferon Long lasting IFNViragen/Valentis Wellferon Interferon Lymphoblastoid GlaxoSmithKlineIFN-αn1 plc, Uxbridge, UK Omniferon Interferon natural IFN-α ViragenInc., Plantation, FL Pegasys Interferon PEGylated IFN-α2a F. Hoffmann-LaRoche LTD, Basel, Switzerland Pegasys and Interferon PEGylated IFN-α2a/Maxim Ceplene immune modulator Pharmaceuticals Inc., San Diego, CAPegasys and Interferon PEGylated IFN- F. Hoffmann-La Ribavirinα2a/ribavirin Roche LTD, Basel, Switzerland PEG-Intron InterferonPEGylated IFN-α2b Schering-Plough Corporation, Kenilworth, NJPEG-Intron/ Interferon PEGylated IFN- Schering-Plough Ribavirinα2b/ribavirin Corporation, Kenilworth, NJ IP-501 Liver protectionAntifibrotic Indevus Pharmaceuticals Inc., Lexington, MA IDN-6556 Liverprotection Caspase inhibitor Idun Pharmaceuticals Inc., San Diego, CAITMN-191 (R- Antiviral Serine protease InterMune 7227) inhibitorPharmaceuticals Inc., Brisbane, CA GL-59728 Antiviral NS5B replicaseGenelabs inhibitor ANA-971 Antiviral TLR-7 agonist Anadys BoceprevirAntiviral Serine protease Schering Plough inhibitor TMS-435 AntiviralSerine protease Tibotec BVBA, inhibitor Mechelen, Belgium BI-201335Antiviral Serine protease Boehringer inhibitor Ingelheim Pharma KG,Ingelheim, Germany MK-7009 Antiviral Serine protease Merck inhibitorPF-00868554 Antiviral Replicase inhibitor Pfizer ANA598 AntiviralNon-Nucleoside Anadys NS5B polymerase Pharmaceuticals, inhibitor Inc.,San Diego, CA, USA IDX375 Antiviral Non-Nucleoside IdenixPharmaceuticals, replicase inhibitor Cambridge, MA, USA BILB 1941Antiviral NS5B polymerase Boehringer inhibitor Ingelheim Canada Ltd R&D,Laval, QC, Canada PSI-7851 Antiviral Nucleoside Pharmasset, polymerasePrinceton, NJ, USA inhibitor VCH-759 Antiviral NS5B polymerase ViroChemPharma inhibitor VCH-916 Antiviral NS5B polymerase ViroChem Pharmainhibitor GS-9190 Antiviral NS5B polymerase Gilead inhibitorPeg-interferon Antiviral Interferon ZymoGenetics/ lamda Bristol-MyersSquibb

The compounds of the present disclosure may also be used as laboratoryreagents. Compounds may be instrumental in providing research tools fordesigning of viral replication assays, validation of animal assaysystems and structural biology studies to further enhance knowledge ofthe HCV disease mechanisms. Further, the compounds of the presentdisclosure are useful in establishing or determining the binding site ofother antiviral compounds, for example, by competitive inhibition.

The compounds of this disclosure may also be used to treat or preventviral contamination of materials and therefore reduce the risk of viralinfection of laboratory or medical personnel or patients who come incontact with such materials, e.g., blood, tissue, surgical instrumentsand garments, laboratory instruments and garments, and blood collectionor transfusion apparatuses and materials.

This disclosure is intended to encompass compounds having Formula (I)when prepared by synthetic processes or by metabolic processes includingthose occurring in the human or animal body (in vivo) or processesoccurring in vitro.

The abbreviations used in the present application, includingparticularly in the illustrative examples which follow, are well-knownto those skilled in the art. Some of the abbreviations used are asfollows:

-   Et ethyl;-   t-Bu tert-butyl;-   iPr isopropyl;-   min minutes;-   rt or RT room temperature or retention time (context will dictate);-   TFA trifluoroacetic acid;-   h or hr hours;-   DMSO dimethylsulfoxide;-   DME dimethyl ether;-   LDA lithium diisopropylamide;-   NBS N-bromosuccinimide;-   SEM-Cl 2-(trimethylsilyl)ethoxymethyl chloride;-   TBAF tetrabutylammonium fluoride;-   HATU O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium    hexafluorophosphate;-   iPr₂EtN diisopropylethylamine;-   DIEA diisopropylethylamine;-   DIPEA diisopropylethylamine;-   Hunig's diisopropylethylamine;-   Boc or BOC tert-butoxycarbonyl;-   DMAP 4-dimethylaminopyridine;-   HCl hydrochloric acid;-   Na₂SO₄ sodium sulfate;-   MgSO₄ magnesium sulfate,-   PdCl₂(PPh₃)₂ bis(triphenylphosphine)palladium(II)dichloride; MCX    cartridge Waters OASIS® MCX LP extraction cartridge.

The compounds and processes of the present disclosure will be betterunderstood in connection with the following synthetic schemes whichillustrate the methods by which the compounds of the present disclosuremay be prepared. Starting materials can be obtained from commercialsources or prepared by well-established literature methods known tothose of ordinary skill in the art. It will be readily apparent to oneof ordinary skill in the art that the compounds defined above can besynthesized by substitution of the appropriate reactants and agents inthe syntheses shown below. It will also be readily apparent to oneskilled in the art that the selective protection and deprotection steps,as well as the order of the steps themselves, can be carried out invarying order, depending on the nature of the variables to successfullycomplete the syntheses below. The variables are as defined above unlessotherwise noted below.

Scheme 1: Substituted Phenylglycine Derivatives

Substituted phenylglycine derivatives can be prepared by a number ofmethods shown below. Phenylglycine t-butyl ester can be reductivelyalkylated (pathyway A) with an appropriate aldehyde and a reductant suchas sodium cyanoborohydride in acidic medium. Hydrolysis of the t-butylester can be accomplished with strong acid such as HCl ortrifluoroacetic acid. Alternatively, phenylglycine can be alkylated withan alkyl halide such as ethyl iodide and a base such as sodiumbicarbonate or potassium carbonate (pathway B). Pathway C illustratesreductive alkylation of phenylglycine as in pathway A followed by asecond reductive alkylation with an alternate aldehyde such asformaldehyde in the presence of a reducing agent and acid. Pathway Dillustrates the synthesis of substituted phenylglycines via thecorresponding mandelic acid analogs. Conversion of the secondary alcoholto a competent leaving group can be accomplished with p-toluensulfonylchloride. Displacement of the tosylate group with an appropriate aminefollowed by reductive removal of the benzyl ester can providesubstituted phenylglycine derivatives. In pathway E a racemicsubstituted phenylglycine derivative is resolved by esterification withan enantiomerically pure chiral auxiliary such as but not limited to(+)-1-phenylethanol, (−)-1-phenylethanol, an Evan's oxazolidinone, orenantiomerically pure pantolactone. Separation of the diastereomers isaccomplished via chromatography (silica gel, HPLC, crystallization, etc)followed by removal of the chiral auxiliary providing enantiomericallypure phenylglycine derivatives. Pathway H illustrates a syntheticsequence which intersects with pathway E wherein the aforementionedchiral auxiliary is installed prior to amine addition. Alternatively, anester of an arylacetic acid can be brominated with a source of bromoniumion such as bromine, N-bromosuccinimide, or CBr₄. The resultant benzylicbromide can be displaced with a variety of mono- or disubstituted aminesin the presence of a tertiary amine base such as triethylamine orHunig's base. Hydrolysis of the methyl ester via treatment with lithiumhydroxide at low temperature or 6N HCl at elevated temperature providesthe substituted phenylglycine derivatives. Another method is shown inpathway G. Glycine analogs can be derivatized with a variety of arylhalides in the presence of a source of palladium (0) such as palladiumbis(tributylphosphine) and base such as potassium phosphate. Theresultant ester can then be hydrolyzed by treatment with base or acid.It should be understood that other well known methods to preparephenylglycine derivatives exist in the art and can be amended to providethe desired compounds in this description. It should also be understoodthat the final phenylglycine derivatives can be purified to enantiomericpurity greater than 98% ee via preparative HPLC.

In another embodiment of the present disclosure, acylated phenylglycinederivatives may be prepared as illustrated below. Phenylglycinederivatives wherein the carboxylic acid is protected as an easilyremoved ester, may be acylated with an acid chloride in the presence ofa base such as triethylamine to provide the corresponding amides(pathway A). Pathway B illustrates the acylation of the startingphenylglycine derivative with an appropriate chloroformate while pathwayC shows reaction with an appropriate isocyanate or carbamoyl chloride.Each of the three intermediates shown in pathways A-C may be deprotectedby methods known by those skilled in the art (ie; treatment of thet-butyl ester with strong base such as HCl or trifluoroacetic acid).

Amino-substituted phenylacetic acids may be prepared by treatment of achloromethylphenylacetic acid with an excess of an amine

Synthesis of Common Caps

Compound Analysis Conditions:

Purity assessment and low resolution mass analysis were conducted on aShimadzu LC system coupled with Waters Micromass ZQ MS system. It shouldbe noted that retention times may vary slightly between machines.Additional LC conditions applicable to the current section, unless notedotherwise.

Cond.-MS-W1

Column = XTERRA 3.0 × 50 mm S7 Start % B =  0 Final % B = 100 Gradienttime = 2 min Stop time = 3 min Flow Rate = 5 mL/min Wavelength = 220 nmSolvent A = 0.1% TFA in 10% methanol/90% H₂O Solvent B = 0.1% TFA in 90%methanol/10% H₂OCond.-MS-W2

Column = XTERRA 3.0 × 50 mm S7 Start % B =  0 Final % B = 100 Gradienttime = 3 min Stop time = 4 min Flow Rate = 4 mL/min Wavelength = 220 nmSolvent A = 0.1% TFA in 10% methanol/90% H₂O Solvent B = 0.1% TFA in 90%methanol/10% H₂OCond.-MS-W5

Column = XTERRA 3.0 × 50 mm S7 Start % B =  0 Final % B = 30 Gradienttime = 2 min Stop time = 3 min Flow Rate = 5 mL/min Wavelength = 220 nmSolvent A = 0.1% TFA in 10% methanol/90% H₂O Solvent B = 0.1% TFA in 90%methanol/10% H₂OCond.-D1

Column = XTERRA C18 3.0 × 50 mm S7 Start % B =  0 Final % B = 100Gradient time = 3 min Stop time = 4 min Flow Rate = 4 mL/min Wavelength= 220 nm Solvent A = 0.1% TFA in 10% methanol/90% H₂O Solvent B = 0.1%TFA in 90% methanol/10% H₂OCond.-D2

Column = Phenomenex-Luna 4.6 × 50 mm S10 Start % B =  0 Final % B = 100Gradient time = 3 min Stop time = 4 min Flow Rate = 4 mL/min Wavelength= 220 nm Solvent A = 0.1% TFA in 10% methanol/90% H₂O Solvent B = 0.1%TFA in 90% methanol/10% H₂OCond.-MD1

Column = XTERRA 4.6 × 50 mm S5 Start % B =  0 Final % B = 100 Gradienttime = 3 min Stop time = 4 min Flow Rate = 4 mL/min Wavelength = 220 nmSolvent A = 0.1% TFA in 10% methanol/90% H₂O Solvent B = 0.1% TFA in 90%methanol/10% H₂OCond.-M3

Column = XTERRA C18 3.0 × 50 mm S7 Start % B =  0 Final % B = 40Gradient time = 2 min Stop time = 3 min Flow Rate = 5 mL/min Wavelength= 220 nm Solvent A = 0.1% TFA in 10% methanol/90% H₂O Solvent B = 0.1%TFA in 90% methanol/10% H₂OCondition OL1

Column = Phenomenex-Luna 3.0 × 50 mm S10 Start % B =  0 Final % B = 100Gradient time = 4 min Stop time = 5 min Flow Rate = 4 mL/min Wavelength= 220 nm Solvent A = 0.1% TFA in 10% methanol/90% H₂O Solvent B = 0.1%TFA in 90% methanol/10% H₂OCondition OL2

Column = Phenomenex-Luna 50 × 2 mm 3 u Start % B =  0 Final % B = 100Gradient time = 4 min Stop time = 5 min Flow Rate = 0.8 mL/min Oven Temp= 40° C. Wavelength = 220 nm Solvent A = 0.1% TFA in 10%Acetonitrile/90% H₂O Solvent B = 0.1% TFA in 90% Acetonitrile/10% H₂OCondition I

Column = Phenomenex-Luna 3.0 × 50 mm S10 Start % B =  0 Final % B = 100Gradient time = 2 min Stop time = 3 min Flow Rate = 4 mL/min Wavelength= 220 nm Solvent A = 0.1% TFA in 10% methanol/90% H₂O Solvent B = 0.1%TFA in 90% methanol/10% H₂OCondition II

Column = Phenomenex-Luna 4.6 × 50 mm S10 Start % B =  0 Final % B = 100Gradient time = 2 min Stop time = 3 min Flow Rate = 5 mL/min Wavelength= 220 nm Solvent A = 0.1% TFA in 10% methanol/90% H₂O Solvent B = 0.1%TFA in 90% methanol/10% H₂OCondition III

Column = XTERRA C18 3.0 × 50 mm S7 Start % B =  0 Final % B = 100Gradient time = 3 min Stop time = 4 min Flow Rate = 4 mL/min Wavelength= 220 nm Solvent A = 0.1% TFA in 10% methanol/90% H₂O Solvent B = 0.1%TFA in 90% methanol/10% H₂O

Cap-1

A suspension of 10% Pd/C (2.0 g) in methanol (10 mL) was added to amixture of (R)-2-phenylglycine (10 g, 66.2 mmol), formaldehyde (33 mL of37% wt. in water), 1N HCl (30 mL) and methanol (30 mL), and exposed toH₂ (60 psi) for 3 hours. The reaction mixture was filtered throughdiatomaceous earth (Celite®), and the filtrate was concentrated invacuo. The resulting crude material was recrystallized from isopropanolto provide the HCl salt of Cap-1 as a white needle (4.0 g). Opticalrotation: −117.1° [c=9.95 mg/mL in H₂O; λ=589 nm]. ¹H NMR (DMSO-d₆,δ=2.5 ppm, 500 MHz): δ 7.43-7.34 (m, 5H), 4.14 (s, 1H), 2.43 (s, 6H); LC(Cond. I): RT=0.25; LC/MS: Anal. Calcd. for [M+H]⁺ C₁₀H₁₄NO₂ 180.10.found 180.17; HRMS: Anal. Calcd. for [M+H]⁺ C₁₀H₁₄NO₂ 180.1025. found180.1017.

Cap-2

NaBH₃CN (6.22 g, 94 mmol) was added in portions over a few minutes to acooled (ice/water) mixture of (R)-2-Phenylglycine (6.02 g, 39.8 mmol)and methanol (100 mL), and stirred for 5 minutes. Acetaldehyde (10 mL)was added dropwise over 10 minutes and stirring was continued at thesame cooled temperature for 45 minutes and at ambient temperature for˜6.5 hours. The reaction mixture was cooled back with ice-water bath,treated with water (3 mL) and then quenched with a dropwise addition ofconcentrated HCl over ˜45 minutes until the pH of the mixture was˜1.5-2.0. The cooling bath was removed and the stirring was continuedwhile adding concentrated HCl in order to maintain the pH of the mixturearound 1.5-2.0. The reaction mixture was stirred overnight, filtered toremove the white suspension, and the filtrate was concentrated in vacuo.The crude material was recrystallized from ethanol to afford the HClsalt of Cap-2 as a shining white solid in two crops (crop-1: 4.16 g;crop-2: 2.19 g). ¹H NMR (DMSO-d₆, δ=2.5 ppm, 400 MHz): 10.44 (1.00, brs, 1H), 7.66 (m, 2H), 7.51 (m, 3H), 5.30 (s, 1H), 3.15 (br m, 2H), 2.98(br m, 2H), 1.20 (app br s, 6H). Crop-1: [α]²⁵ −102.21° (c=0.357, H₂O);crop-2: [α]²⁵ −99.7° (c=0.357, H₂O). LC (Cond. I): RT=0.43 min; LC/MS:Anal. Calcd. for [M+H]⁺ C₁₂H₁₈NO₂: 208.13. found 208.26.

Cap-3

Acetaldehyde (5.0 mL, 89.1 mmol) and a suspension of 10% Pd/C (720 mg)in methanol/H₂O (4 mL/1 mL) was sequentially added to a cooled (˜15° C.)mixture of (R)-2-phenylglycine (3.096 g, 20.48 mmol), 1N HCl (30 mL) andmethanol (40 mL). The cooling bath was removed and the reaction mixturewas stirred under a balloon of H₂ for 17 hours. An additionalacetaldehyde (10 mL, 178.2 mmol) was added and stirring continued underH₂ atmosphere for 24 hours [Note: the supply of H₂ was replenished asneeded throughout the reaction]. The reaction mixture was filteredthrough diatomaceous earth (Celite®), and the filtrate was concentratedin vacuo. The resulting crude material was recrystallized fromisopropanol to provide the HCl salt of (R)-2-(ethylamino)-2-phenylaceticacid as a shining white solid (2.846 g). ¹H NMR (DMSO-d₆, δ=2.5 ppm, 400MHz): δ 14.15 (br s, 1H), 9.55 (br s, 2H), 7.55-7.48 (m, 5H), 2.88 (brm, 1H), 2.73 (br m, 1H), 1.20 (app t, J=7.2, 3H). LC (Cond. I): RT=0.39min; >95% homogeneity index; LC/MS: Anal. Calcd. for [M+H]⁺ C₁₀H₁₄NO₂:180.10. found 180.18.

A suspension of 10% Pd/C (536 mg) in methanol/H₂O (3 mL/1 mL) was addedto a mixture of (R)-2-(ethylamino)-2-phenylacetic acid/HCl (1.492 g,6.918 mmol), formaldehyde (20 mL of 37% wt. in water), 1N HCl (20 mL)and methanol (23 mL). The reaction mixture was stirred under a balloonof H₂ for ˜72 hours, where the H₂ supply was replenished as needed. Thereaction mixture was filtered through diatomaceous earth (Celite®) andthe filtrate was concentrated in vacuo. The resulting crude material wasrecrystallized from isopropanol (50 mL) to provide the HCl salt of Cap-3as a white solid (985 mg). ¹H NMR (DMSO-d₆, δ=2.5 ppm, 400 MHz): δ 10.48(br s, 1H), 7.59-7.51 (m, 5H), 5.26 (s, 1H), 3.08 (app br s, 2H), 2.65(br s, 3H), 1.24 (br m, 3H). LC (Cond. I): RT=0.39 min; >95% homogeneityindex; LC/MS: Anal. Calcd. for [M+H]⁺ C₁₁H₁₆NO₂: 194.12. found 194.18;HRMS: Anal. Calcd. for [M+H]⁺ C₁₁H₁₆NO₂: 194.1180. found 194.1181.

Cap-4

ClCO₂Me (3.2 mL, 41.4 mmol) was added dropwise to a cooled (ice/water)THF (410 mL) semi-solution of (R)-tert-butyl 2-amino-2-phenylacetate/HCl(9.877 g, 40.52 mmol) and diisopropylethylamine (14.2 mL, 81.52 mmol)over 6 min, and stirred at similar temperature for 5.5 hours. Thevolatile component was removed in vacuo, and the residue was partitionedbetween water (100 mL) and ethyl acetate (200 mL). The organic layer waswashed with 1N HCl (25 mL) and saturated NaHCO₃ solution (30 mL), dried(MgSO₄), filtered, and concentrated in vacuo. The resultant colorlessoil was triturated from hexanes, filtered and washed with hexanes (100mL) to provide (R)-tert-butyl 2-(methoxycarbonylamino)-2-phenylacetateas a white solid (7.7 g). ¹H NMR (DMSO-d₆, δ=2.5 ppm, 400 MHz): 7.98 (d,J=8.0, 1H), 7.37-7.29 (m, 5H), 5.09 (d, J=8, 1H), 3.56 (s, 3H), 1.33 (s,9H). LC (Cond. I): RT=1.53 min; ˜90% homogeneity index; LC/MS: Anal.Calcd. for [M+Na]⁺ C₁₄H₁₉NNaO₄: 288.12. found 288.15.

TFA (16 mL) was added dropwise to a cooled (ice/water) CH₂Cl₂ (160 mL)solution of the above product over 7 minutes, and the cooling bath wasremoved and the reaction mixture was stirred for 20 hours. Since thedeprotection was still not complete, an additional TFA (1.0 mL) wasadded and stirring continued for an additional 2 hours. The volatilecomponent was removed in vacuo, and the resulting oil residue wastreated with diethyl ether (15 mL) and hexanes (12 mL) to provide aprecipitate. The precipitate was filtered and washed with diethylether/hexanes (˜1:3 ratio; 30 mL) and dried in vacuo to provide Cap-4 asa fluffy white solid (5.57 g). Optical rotation: −176.9° [c=3.7 mg/mL inH₂O; λ=589 nm]. ¹H NMR (DMSO-d₆, δ=2.5 ppm, 400 MHz): δ 12.84 (br s,1H), 7.96 (d, J=8.3, 1H), 7.41-7.29 (m, 5H), 5.14 (d, J=8.3, 1H), 3.55(s, 3H). LC (Cond. I): RT=1.01 min; >95% homogeneity index; LC/MS: Anal.Calcd. for [M+H]⁺ C₁₀H₁₂NO₄ 210.08. found 210.17; HRMS: Anal. Calcd. for[M+H]⁺ C₁₀H₁₂NO₄ 210.0766. found 210.0756.

Cap-5

A mixture of (R)-2-phenylglycine (1.0 g, 6.62 mmol), 1,4-dibromobutane(1.57 g, 7.27 mmol) and Na₂CO₃ (2.10 g, 19.8 mmol) in ethanol (40 mL)was heated at 100° C. for 21 hours. The reaction mixture was cooled toambient temperature and filtered, and the filtrate was concentrated invacuo. The residue was dissolved in ethanol and acidified with 1N HCl topH 3-4, and the volatile component was removed in vacuo. The resultingcrude material was purified by a reverse phase HPLC (water/methanol/TFA)to provide the TFA salt of Cap-5 as a semi-viscous white foam (1.0 g).¹H NMR (DMSO-d₆, δ=2.5, 500 MHz) δ 10.68 (br s, 1H), 7.51 (m, 5H), 5.23(s, 1H), 3.34 (app br s, 2H), 3.05 (app br s, 2H), 1.95 (app br s, 4H);RT=0.30 minutes (Cond. I); >98% homogeneity index; LC/MS: Anal. Calcd.for [M+H]⁺ C₁₂H₁₆NO₂: 206.12. found 206.25.

Cap-6

The TFA salt of Cap-6 was synthesized from (R)-2-phenylglycine and1-bromo-2-(2-bromoethoxy)ethane by using the method of preparation ofCap-5. ¹H NMR (DMSO-d₆, δ=2.5, 500 MHz) δ 12.20 (br s, 1H), 7.50 (m,5H), 4.92 (s, 1H), 3.78 (app br s, 4H), 3.08 (app br s, 2H), 2.81 (appbr s, 2H); RT=0.32 minutes (Cond. I); >98%; LC/MS: Anal. Calcd. for[M+H]⁺ C₁₂H₁₆NO₃: 222.11. found 222.20; HRMS: Anal. Calcd. for [M+H]⁺C₁₂H₁₆NO₃: 222.1130. found 222.1121.

Cap-7

A CH₂Cl₂ (200 mL) solution of p-toluenesulfonyl chloride (8.65 g, 45.4mmol) was added dropwise to a cooled (−5° C.) CH₂Cl₂ (200 mL) solutionof (S)-benzyl 2-hydroxy-2-phenylacetate (10.0 g, 41.3 mmol),triethylamine (5.75 mL, 41.3 mmol) and 4-dimethylaminopyridine (0.504 g,4.13 mmol), while maintaining the temperature between −5° C. and 0° C.The reaction was stirred at 0° C. for 9 hours, and then stored in afreezer (−25° C.) for 14 hours. It was allowed to thaw to ambienttemperature and washed with water (200 mL), 1N HCl (100 mL) and brine(100 mL), dried (MgSO₄), filtered, and concentrated in vacuo to providebenzyl 2-phenyl-2-(tosyloxy)acetate as a viscous oil which solidifiedupon standing (16.5 g). The chiral integrity of the product was notchecked and that product was used for the next step without furtherpurification. ¹H NMR (DMSO-d₆, δ=2.5, 500 MHz) δ 7.78 (d, J=8.6, 2H),7.43-7.29 (m, 10H), 7.20 (m, 2H), 6.12 (s, 1H), 5.16 (d, J=12.5, 1H),5.10 (d, J=12.5, 1H), 2.39 (s, 3H). RT=3.00 (Cond. III); >90%homogeneity index; LC/MS: Anal. Calcd. for [M+H]⁺ C₂₂H₂₀NaO₅S: 419.09.found 419.04.

A THF (75 mL) solution of benzyl 2-phenyl-2-(tosyloxy)acetate (6.0 g,15.1 mmol), 1-methylpiperazine (3.36 mL, 30.3 mmol) andN,N-diisopropylethylamine (13.2 mL, 75.8 mmol) was heated at 65° C. for7 hours. The reaction was allowed to cool to ambient temperature and thevolatile component was removed in vacuo. The residue was partitionedbetween ethylacetate and water, and the organic layer was washed withwater and brine, dried (MgSO₄), filtered, and concentrated in vacuo. Theresulting crude material was purified by flash chromatography (silicagel, ethyl acetate) to provide benzyl2-(4-methylpiperazin-1-yl)-2-phenylacetate as an orangish-brown viscousoil (4.56 g). Chiral HPLC analysis (Chiralcel OD-H) indicated that thesample is a mixture of enantiomers in a 38.2 to 58.7 ratio. Theseparation of the enantiomers were effected as follow: the product wasdissolved in 120 mL of ethanol/heptane (1:1) and injected (5mL/injection) on chiral HPLC column (Chiracel OJ, 5 cm ID×50 cm L, 20μm) eluting with 85:15 Heptane/ethanol at 75 mL/min, and monitored at220 nm. Enantiomer-1 (1.474 g) and enantiomer-2 (2.2149 g) wereretrieved as viscous oil. ¹H NMR (CDCl₃, δ=7.26, 500 MHz) 7.44-7.40 (m,2H), 7.33-7.24 (m, 6H), 7.21-7.16 (m, 2H), 5.13 (d, J=12.5, 1H), 5.08(d, J=12.5, 1H), 4.02 (s, 1H), 2.65-2.38 (app br s, 8H), 2.25 (s, 3H).RT=2.10 (Cond. III); >98% homogeneity index; LC/MS: Anal. Calcd. for[M+H]⁺ C₂₀H₂₅N₂O₂: 325.19. found 325.20.

A methanol (10 mL) solution of either enantiomer of benzyl2-(4-methylpiperazin-1-yl)-2-phenylacetate (1.0 g, 3.1 mmol) was addedto a suspension of 10% Pd/C (120 mg) in methanol (5.0 mL). The reactionmixture was exposed to a balloon of hydrogen, under a carefulmonitoring, for <50 minutes. Immediately after the completion of thereaction, the catalyst was filtered through diatomaceous earth (Celite®)and the filtrate was concentrated in vacuo to provide Cap-7,contaminated with phenylacetic acid as a tan foam (867.6 mg; mass isabove the theoretical yield). The product was used for the next stepwithout further purification. ¹H NMR (DMSO-d₆, δ=2.5, 500 MHz) δ7.44-7.37 (m, 2H), 7.37-7.24 (m, 3H), 3.92 (s, 1H), 2.63-2.48 (app. brs, 2H), 2.48-2.32 (m, 6H), 2.19 (s, 3H); RT=0.31 (Cond. II); >90%homogeneity index; LC/MS: Anal. Calcd. for [M+H]⁺ C₁₃H₁₉N₂O₂: 235.14.found 235.15; HRMS: Anal. Calcd. for [M+H]⁺ C₁₃H₁₉N₂O₂: 235.1447. found235.1440.

The synthesis of Cap-8 and Cap-9 was conducted according to thesynthesis of Cap-7 by using appropriate amines for the SN₂ displacementstep (i.e., 4-hydroxypiperidine for Cap-8 and (S)-3-fluoropyrrolidinefor Cap-9) and modified conditions for the separation of the respectivestereoisomeric intermedites, as described below.

Cap-8

The enantiomeric separation of the intermediate benzyl2-(4-hydroxypiperidin-1-yl)-2-phenyl acetate was effected by employingthe following conditions: the compound (500 mg) was dissolved inethanol/heptane (5 mL/45 mL). The resulting solution was injected (5mL/injection) on a chiral HPLC column (Chiracel OJ, 2 cm ID×25 cm L, 10μm) eluting with 80:20 heptane/ethanol at 10 mL/min, monitored at 220nm, to provide 186.3 mg of enantiomer-1 and 209.1 mg of enantiomer-2 aslight-yellow viscous oils. These benzyl ester was hydrogenolysedaccording to the preparation of Cap-7 to provide Cap-8: ¹H NMR (DMSO-d₆,δ=2.5, 500 MHz) 7.40 (d, J=7, 2H), 7.28-7.20 (m, 3H), 3.78 (s 1H), 3.46(m, 1H), 2.93 (m, 1H), 2.62 (m, 1H), 2.20 (m, 2H), 1.70 (m, 2H), 1.42(m, 2H). RT=0.28 (Cond. II); >98% homogeneity index; LC/MS: Anal. Calcd.for [M+H]⁺ C₁₃H₁₈NO₃: 236.13. found 236.07; HRMS: Calcd. for [M+H]⁺C₁₃H₁₈NO₃: 236.1287. found 236.1283.

Cap-9

The diastereomeric separation of the intermediate benzyl2-((S)-3-fluoropyrrolidin-1-yl)-2-phenylacetate was effected byemploying the following conditions: the ester (220 mg) was separated ona chiral HPLC column (Chiracel OJ-H, 0.46 cm ID×25 cm L, 5 μm) elutingwith 95% CO₂/5% methanol with 0.1% TFA, at 10 bar pressure, 70 mL/minflow rate, and a temperature of 35° C. The HPLC elute for the respectivestereoisomers was concentrated, and the residue was dissolved in CH₂Cl₂(20 mL) and washed with an aqueous medium (10 mL water+1 mL saturatedNaHCO₃ solution). The organic phase was dried (MgSO₄), filtered, andconcentrated in vacuo to provide 92.5 mg of fraction-1 and 59.6 mg offraction-2. These benzyl esters were hydrogenolysed according to thepreparation of Cap-7 to prepare Caps 9a and 9b. Cap-9a (diastereomer-1;the sample is a TFA salt as a result of purification on a reverse phaseHPLC using H₂O/methanol/TFA solvent): ¹H NMR (DMSO-d₆, δ=2.5, 400 MHz)7.55-7.48 (m, 5H), 5.38 (d of m, J=53.7, 1H), 5.09 (br s, 1H), 3.84-2.82(br m, 4H), 2.31-2.09 (m, 2H). RT=0.42 (Cond. I); >95% homogeneityindex; LC/MS: Anal. Calcd. for [M+H]⁺ C₁₂H₁₅FNO₂: 224.11. found 224.14;Cap-9b (diastereomer-2): ¹H NMR (DMSO-d₆, δ=2.5, 400 MHz) 7.43-7.21 (m,5H), 5.19 (d of m, J=55.9, 1H), 3.97 (s, 1H), 2.95-2.43 (m, 4H),2.19-1.78 (m, 2H). RT=0.44 (Cond. I); LC/MS: Anal. Calcd. for [M+H]⁺C₁₂H₁₅FNO₂: 224.11. found 224.14.

Cap-10

To a solution of D-proline (2.0 g, 17 mmol) and formaldehyde (2.0 mL of37% wt. in H₂O) in methanol (15 mL) was added a suspension of 10% Pd/C(500 mg) in methanol (5 mL). The mixture was stirred under a balloon ofhydrogen for 23 hours. The reaction mixture was filtered throughdiatomaceous earth (Celite®) and concentrated in vacuo to provide Cap-10as an off-white solid (2.15 g). ¹H NMR (DMSO-d₆, δ=2.5, 500 MHz) 3.42(m, 1H), 3.37 (dd, J=9.4, 6.1, 1H), 2.85-2.78 (m, 1H), 2.66 (s, 3H),2.21-2.13 (m, 1H), 1.93-1.84 (m, 2H), 1.75-1.66 (m, 1H). RT=0.28 (Cond.II); >98% homogeneity index; LC/MS: Anal. Calcd. for [M+H]⁺ C₆H₁₂NO₂:130.09. found 129.96.

Cap-11

A mixture of (2S,4R)-4-fluoropyrrolidine-2-carboxylic acid (0.50 g, 3.8mmol), formaldehyde (0.5 mL of 37% wt. in H₂O), 12 N HCl (0.25 mL) and10% Pd/C (50 mg) in methanol (20 mL) was stirred under a balloon ofhydrogen for 19 hours. The reaction mixture was filtered throughdiatomaceous earth (Celite®) and the filtrate was concentrated in vacuo.The residue was recrystallized from isopropanol to provide the HCl saltof Cap-11 as a white solid (337.7 mg). ¹H NMR (DMSO-d₆, δ=2.5, 500 MHz)5.39 (d m, J=53.7, 1H), 4.30 (m, 1H), 3.90 (ddd, J=31.5, 13.5, 4.5, 1H),3.33 (dd, J=25.6, 13.4, 1H), 2.85 (s, 3H), 2.60-2.51 (m, 1H), 2.39-2.26(m, 1H). RT=0.28 (Cond. II); >98% homogeneity index; LC/MS: Anal. Calcd.for [M+H]⁺ C₆H₁₁FNO₂: 148.08. found 148.06.

Cap-12 (Same as Cap 52)

L-Alanine (2.0 g, 22.5 mmol) was dissolved in 10% aqueous sodiumcarbonate solution (50 mL), and a THF (50 mL) solution of methylchloroformate (4.0 mL) was added to it. The reaction mixture was stirredunder ambient conditions for 4.5 hours and concentrated in vacuo. Theresulting white solid was dissolved in water and acidified with 1N HClto a pH ˜2-3. The resulting solutions was extracted with ethyl acetate(3×100 mL), and the combined organic phase was dried (Na₂SO₄), filtered,and concentrated in vacuo to provide a colorless oil (2.58 g). 500 mg ofthis material was purified by a reverse phase HPLC(H₂O/methanol/TFA) toprovide 150 mg of Cap-12 as a colorless oil. ¹H NMR (DMSO-d₆, δ=2.5, 500MHz) 7.44 (d, J=7.3, 0.8H), 7.10 (br s, 0.2H), 3.97 (m, 1H), 3.53 (s,3H), 1.25 (d, J=7.3, 3H).

Cap-13

A mixture of L-alanine (2.5 g, 28 mmol), formaldehyde (8.4 g, 37 wt. %),1N HCl (30 mL) and 10% Pd/C (500 mg) in methanol (30 mL) was stirredunder a hydrogen atmosphere (50 psi) for 5 hours. The reaction mixturewas filtered through diatomaceous earth (Celite®) and the filtrate wasconcentrated in vacuo to provide the HCl salt of Cap-13 as an oil whichsolidified upon standing under vacuum (4.4 g; the mass is abovetheoretical yield). The product was used without further purification.¹H NMR (DMSO-d₆, δ=2.5, 500 MHz) δ 12.1 (br s, 1H), 4.06 (q, J=7.4, 1H),2.76 (s, 6H), 1.46 (d, J=7.3, 3H).

Cap-14

Step 1

A mixture of (R)-(−)-D-phenylglycine tert-butyl ester (3.00 g, 12.3mmol), NaBH₃CN (0.773 g, 12.3 mmol), KOH (0.690 g, 12.3 mmol) and aceticacid (0.352 mL, 6.15 mmol) were stirred in methanol at 0° C. To thismixture was added glutaric dialdehyde (2.23 mL, 12.3 mmol) dropwise over5 minutes. The reaction mixture was stirred as it was allowed to warm toambient temperature and stirring was continued at the same temperaturefor 16 hours. The solvent was subsequently removed and the residue waspartitioned with 10% aqueous NaOH and ethyl acetate. The organic phasewas separated, dried (MgSO₄), filtered and concentrated to dryness toprovide a clear oil. This material was purified by reverse-phasepreparative HPLC (Primesphere C-18, 30×100 mm; CH₃CN—H₂O-0.1% TFA) togive the intermediate ester (2.70 g, 56%) as a clear oil. ¹H NMR (400MHz, CDCl₃) δ 7.53-7.44 (m, 3H), 7.40-7.37 (m, 2H), 3.87 (d, J=10.9 Hz,1H), 3.59 (d, J=10.9 Hz, 1H), 2.99 (t, J=11.2 Hz, 1H), 2.59 (t, J=11.4Hz, 1H), 2.07-2.02 (m, 2H), 1.82 (d, J=1.82 Hz, 3H), 1.40 (s, 9H).LC/MS: Anal. Calcd. for C₁₇H₂₅NO₂: 275. found: 276 (M+H)⁺.

Step 2

To a stirred solution of the intermediate ester (1.12 g, 2.88 mmol) indichloromethane (10 mL) was added TFA (3 mL). The reaction mixture wasstirred at ambient temperature for 4 hours and then it was concentratedto dryness to give a light yellow oil. The oil was purified usingreverse-phase preparative HPLC (Primesphere C-18, 30×100 mm;CH₃CN—H₂O-0.1% TFA). The appropriate fractions were combined andconcentrated to dryness in vacuo. The residue was then dissolved in aminimum amount of methanol and applied to applied to MCX LP extractioncartridges (2×6 g). The cartridges were rinsed with methanol (40 mL) andthen the desired compound was eluted using 2M ammonia in methanol (50mL). Product-containing fractions were combined and concentrated and theresidue was taken up in water. Lyophilization of this solution providedthe title compound (0.492 g, 78%) as a light yellow solid. ¹H NMR(DMSO-d₆) δ 7.50 (s, 5H), 5.13 (s, 1H), 3.09 (br s, 2H), 2.92-2.89 (m,2H), 1.74 (m, 4H), 1.48 (br s, 2H). LC/MS: Anal. Calcd. for C₁₃H₁₂NO₂:219. found: 220 (M+H)⁺.

Cap-15

Step 1

(S)-1-Phenylethyl 2-bromo-2-phenylacetate: To a mixture ofα-bromophenylacetic acid (10.75 g, 0.050 mol), (S)-(−)-1-phenylethanol(7.94 g, 0.065 mol) and DMAP (0.61 g, 5.0 mmol) in dry dichloromethane(100 mL) was added solid EDCI (12.46 g, 0.065 mol) all at once. Theresulting solution was stirred at room temperature under Ar for 18 hoursand then it was diluted with ethyl acetate, washed (H₂O×2, brine), dried(Na₂SO₄), filtered, and concentrated to give a pale yellow oil. Flashchromatography (SiO₂/hexane-ethyl acetate, 4:1) of this oil provided thetitle compound (11.64 g, 73%) as a white solid. ¹H NMR (400 MHz, CDCl₃)δ 7.53-7.17 (m, 10H), 5.95 (q, J=6.6 Hz, 0.5H), 5.94 (q, J=6.6 Hz,0.5H), 5.41 (s, 0.5H), 5.39 (s, 0.5H), 1.58 (d, J=6.6 Hz, 1.5H), 1.51(d, J=6.6 Hz, 1.5H).

Step 2

(S)-1-Phenylethyl(R)-2-(4-hydroxy-4-methylpiperidin-1-yl)-2-phenylacetate:To a solution of (S)-1-phenylethyl 2-bromo-2-phenylacetate (0.464 g,1.45 mmol) in THF (8 mL) was added triethylamine (0.61 mL, 4.35 mmol),followed by tetrabutylammonium iodide (0.215 g, 0.58 mmol). The reactionmixture was stirred at room temperature for 5 minutes and then asolution of 4-methyl-4-hydroxypiperidine (0.251 g, 2.18 mmol) in THF (2mL) was added. The mixture was stirred for 1 hour at room temperatureand then it was heated at 55-60° C. (oil bath temperature) for 4 hours.The cooled reaction mixture was then diluted with ethyl acetate (30 mL),washed (H₂O×2, brine), dried (MgSO₄), filtered and concentrated. Theresidue was purified by silica gel chromatography (0-60% ethylacetate-hexane) to provide first the (S,R)-isomer of the title compound(0.306 g, 60%) as a white solid and then the corresponding (S,S)-isomer(0.120 g, 23%), also as a white solid. (S,R)-isomer: ¹H NMR (CD₃OD) δ7.51-7.45 (m, 2H), 7.41-7.25 (m, 8H), 5.85 (q, J=6.6 Hz, 1H), 4.05 (s,1H), 2.56-2.45 (m, 2H), 2.41-2.29 (m, 2H), 1.71-1.49 (m, 4H), 1.38 (d,J=6.6 Hz, 3H), 1.18 (s, 3H). LCMS: Anal. Calcd. for C₂₂H₂₇NO₃: 353.found: 354 (M+H)⁺. (S,S)-isomer: ¹H NMR (CD₃OD) δ 7.41-7.30 (m, 5H),7.20-7.14 (m, 3H), 7.06-7.00 (m, 2H), 5.85 (q, J=6.6 Hz, 1H), 4.06 (s,1H), 2.70-2.60 (m, 1H), 2.51 (dt, J=6.6, 3.3 Hz, 1H), 2.44-2.31 (m, 2H),1.75-1.65 (m, 1H), 1.65-1.54 (m, 3H), 1.50 (d, J=6.8 Hz, 3H), 1.20 (s,3H). LCMS: Anal. Calcd. for C₂₂H₂₇NO₃: 353. found: 354 (M+H)⁺.

Step 3

(R)-2-(4-Hydroxy-4-methylpiperidin-1-yl)-2-phenylacetic acid: To asolution of(S)-1-phenylethyl(R)-2-(4-hydroxy-4-methylpiperidin-1-yl)-2-phenylacetate(0.185 g, 0.52 mmol) in dichloromethane (3 mL) was added trifluoroaceticacid (1 mL) and the mixture was stirred at room temperature for 2 hours.The volatiles were subsequently removed in vacuo and the residue waspurified by reverse-phase preparative HPLC (Primesphere C-18, 20×100 mm;CH₃CN—H₂O-0.1% TFA) to give the title compound (as TFA salt) as a palebluish solid (0.128 g, 98%). LCMS: Anal. Calcd. for C₁₄H₁₉NO₃: 249.found: 250 (M+H)⁺.

Cap-16

Step 1

(S)-1-Phenylethyl 2-(2-fluorophenyl)acetate: A mixture of2-fluorophenylacetic acid (5.45 g, 35.4 mmol), (S)-1-phenylethanol (5.62g, 46.0 mmol), EDCI (8.82 g, 46.0 mmol) and DMAP (0.561 g, 4.60 mmol) inCH₂Cl₂ (100 mL) was stirred at room temperature for 12 hours. Thesolvent was then concentrated and the residue partitioned with H₂O-ethylacetate. The phases were separated and the aqueous layer back-extractedwith ethyl acetate (2×). The combined organic phases were washed (H₂O,brine), dried (Na₂SO₄), filtered, and concentrated in vacuo. The residuewas purified by silica gel chromatography (Biotage/0-20% ethylacetate-hexane) to provide the title compound as a colorless oil (8.38g, 92%). ¹H NMR (400 MHz, CD₃OD) δ 7.32-7.23 (m, 7H), 7.10-7.04 (m, 2),5.85 (q, J=6.5 Hz, 1H), 3.71 (s, 2H), 1.48 (d, J=6.5 Hz, 3H).

Step 2

(R)—((S)-1-Phenylethyl) 2-(2-fluorophenyl)-2-(piperidin-1-yl)acetate: Toa solution of (S)-1-phenylethyl 2-(2-fluorophenyl)acetate (5.00 g, 19.4mmol) in THF (1200 mL) at 0° C. was added DBU (6.19 g, 40.7 mmol) andthe solution was allowed to warm to room temperature while stirring for30 minutes. The solution was then cooled to −78° C. and a solution ofCBr₄ (13.5 g, 40.7 mmol) in THF (100 mL) was added and the mixture wasallowed to warm to −10° C. and stirred at this temperature for 2 hours.The reaction mixture was quenched with saturated aq. NH₄Cl and thelayers were separated. The aqueous layer was back-extracted with ethylacetate (2×) and the combined organic phases were washed (H₂O, brine),dried (Na₂SO₄), filtered, and concentrated in vacuo. To the residue wasadded piperidine (5.73 mL, 58.1 mmol) and the solution was stirred atroom temperature for 24 hours. The volatiles were then concentrated invacuo and the residue was purified by silica gel chromatography(Biotage/0-30% diethyl ether-hexane) to provide a pure mixture ofdiastereomers (2:1 ratio by ¹H NMR) as a yellow oil (2.07 g, 31%), alongwith unreacted starting material (2.53 g, 51%). Further chromatographyof the diastereomeric mixture (Biotage/0-10% diethyl ether-toluene)provided the title compound as a colorless oil (0.737 g, 11%). ¹H NMR(400 MHz, CD₃OD) δ 7.52 (ddd, J=9.4, 7.6, 1.8 Hz, 1H), 7.33-7.40 (m, 1),7.23-7.23 (m, 4H), 7.02-7.23 (m, 4H), 5.86 (q, J=6.6 Hz, 1H), 4.45 (s,1H), 2.39-2.45 (m, 4H), 1.52-1.58 (m, 4H), 1.40-1.42 (m, 1H), 1.38 (d,J=6.6 Hz, 3H). LCMS: Anal. Calcd. for C₂₁H₂₄FNO₂: 341. found: 342(M+H)⁺.

Step 3

(R)-2-(2-fluorophenyl)-2-(piperidin-1-yl)acetic acid: A mixture of(R)—((S)-1-phenylethyl) 2-(2-fluorophenyl)-2-(piperidin-1-yl)acetate(0.737 g, 2.16 mmol) and 20% Pd(OH)₂/C (0.070 g) in ethanol (30 mL) washydrogenated at room temperature and atmospheric pressure (H₂ balloon)for 2 hours. The solution was then purged with Ar, filtered throughdiatomaceous earth (Celite®), and concentrated in vacuo. This providedthe title compound as a colorless solid (0.503 g, 98%). ¹H NMR (400 MHz,CD₃OD) δ 7.65 (ddd, J=9.1, 7.6, 1.5 Hz, 1H), 7.47-7.53 (m, 1H),7.21-7.30 (m, 2H), 3.07-3.13 (m, 4H), 1.84 (br s, 4H), 1.62 (br s, 2H).LCMS: Anal. Calcd. for C₁₃H₁₆FNO₂: 237. found: 238 (M+H)⁺.

Cap-17

Step 1

(S)-1-Phenylethyl(R)-2-(4-hydroxy-4-phenylpiperidin-1-yl)-2-phenylacetate:To a solution of (S)-1-phenylethyl 2-bromo-2-phenylacetate (1.50 g, 4.70mmol) in THF (25 mL) was added triethylamine (1.31 mL, 9.42 mmol),followed by tetrabutylammonium iodide (0.347 g, 0.94 mmol). The reactionmixture was stirred at room temperature for 5 minutes and then asolution of 4-phenyl-4-hydroxypiperidine (1.00 g, 5.64 mmol) in THF (5mL) was added. The mixture was stirred for 16 hours and then it wasdiluted with ethyl acetate (100 mL), washed (H₂O×2, brine), dried(MgSO₄), filtered and concentrated. The residue was purified on a silicagel column (0-60% ethyl acetate-hexane) to provide an approximately 2:1mixture of diastereomers, as judged by ¹H NMR. Separation of theseisomers was performed using supercritical fluid chromatography(Chiralcel OJ-H, 30×250 mm; 20% ethanol in CO₂ at 35° C.), to give firstthe (R)-isomer of the title compound (0.534 g, 27%) as a yellow oil andthen the corresponding (S)-isomer (0.271 g, 14%), also as a yellow oil.(S,R)-isomer: ¹H NMR (400 MHz, CD₃OD) δ 7.55-7.47 (m, 4H), 7.44-7.25 (m,10H), 7.25-7.17 (m, 1H), 5.88 (q, J=6.6 Hz, 1H), 4.12 (s, 1H), 2.82-2.72(m, 1H), 2.64 (dt, J=11.1, 2.5 Hz, 1H), 2.58-2.52 (m, 1H), 2.40 (dt,J=11.1, 2.5 Hz, 1H), 2.20 (dt, J=12.1, 4.6 Hz, 1H), 2.10 (dt, J=12.1,4.6 Hz, 1H), 1.72-1.57 (m, 2H), 1.53 (d, J=6.5 Hz, 3H). LCMS: Anal.Calcd. for C₂₇H₂₉NO₃: 415. found: 416 (M+H)⁺; (S,S)-isomer: H¹NMR (400MHz, CD₃OD) δ 7.55-7.48 (m, 2H), 7.45-7.39 (m, 2H), 7.38-7.30 (m, 5H),7.25-7.13 (m, 4H), 7.08-7.00 (m, 2H), 5.88 (q, J=6.6 Hz, 1H), 4.12 (s,1H), 2.95-2.85 (m, 1H), 2.68 (dt, J=11.1, 2.5 Hz, 1H), 2.57-2.52 (m,1H), 2.42 (dt, J=11.1, 2.5 Hz, 1H), 2.25 (dt, J=12.1, 4.6 Hz, 1H), 2.12(dt, J=12.1, 4.6 Hz, 1H), 1.73 (dd, J=13.6, 3.0 Hz, 1H), 1.64 (dd,J=13.6, 3.0 Hz, 1H), 1.40 (d, J=6.6 Hz, 3H). LCMS: Anal. Calcd. forC₂₇H₂₉NO₃: 415. found: 416 (M+H)⁺.

The following esters were prepared in similar fashion:

Intermediate-17a

Diastereomer 1: ¹H NMR (500 MHz, DMSO-d₆) δ ppm 1.36 (d, J = 6.41 Hz,3H) 2.23-2.51 (m, 4H) 3.35 (s, 4H) 4.25 (s, 1H) 5.05 (s, 2H) 5.82 (d, J= 6.71 Hz, 1H) 7.15-7.52 (m, 15H). LCMS: Anal. Calcd. for: C₂₈H₃₀N₂O₄458.22; Found: 459.44 (M + H)⁺. Diastereomer 2: ¹HNMR (500 MHz, DMSO-d₆)δ ppm 1.45 (d, J = 6.71 Hz, 3H) 2.27-2.44 (m, 4H) 3.39 (s, 4H) 4.23 (s,1H) 5.06 (s, 2H) 5.83 (d, J = 6.71 Hz, 1H) 7.12 (dd, J = 6.41, 3.05 Hz,2H) 7.19-7.27 (m, 3H) 7.27- 7.44 (m, 10H). LCMS: Anal. Calcd. for:C₂₈H₃₀N₂O₄ 458.22; Found: 459.44 (M + H)⁺. Intermediate-17b

Diasteromer 1: RT ₌ 11.76 minutes (Cond'n II); LCMS: Anal. Calcd. for:C₂₀H₂₂N₂O₃ 338.16 Found: 339.39 (M + H)⁺; Diastercomcr 2: RT =10.05minutes (Cond'n II); LCMS: Anal. Calcd. for: C₂₀H₂₂N₂O₃ 338.16; Found:339.39 (M + H)⁺. Intermediate-17c

Diastereomer 1: T_(R =) 4.55 minutes (Cond'n I); LCMS: Anal. Calcd. for:C₂₁H₂₆N₂O₂ 338.20 Found: 339.45 (M + H)⁺; Diastereomer 2: T_(R =) 6.00minutes (Cond'n I); LCMS: Anal. Calcd. for: C₂₁H₂₆N₂O₂ 338.20 Found:339.45 (M + H)⁺. Intermediate-17d

Diastercomcr 1: RT ₌ 7.19 minutes (Cond'n I); LCMS: Anal. Calcd. for:C₂₇H₂₉NO₂ 399.22 Found: 400.48 (M + H)⁺; Diastercomcr 2: RT = 9.76minutes (Cond'n I); LCMS: Anal. Calcd. for: C₂₇H₂₉NO₂ 399.22 Found:400.48 (M + H)⁺.Chiral SFC Conditions for determining retention timeCondition IColumn: Chiralpak AD-H Column, 4.62×50 mm, 5 μmSolvents: 90% CO2-10% methanol with 0.1% DEATemp: 35° C.Pressure: 150 barFlow rate: 2.0 mL/min.UV monitored @ 220 nmInjection: 1.0 mg/3 mL methanolCondition IIColumn: Chiralcel OD-H Column, 4.62×50 mm, 5 μmSolvents: 90% CO2-10% methanol with 0.1% DEATemp: 35° C.Pressure: 150 barFlow rate: 2.0 mL/min.UV monitored @ 220 nmInjection: 1.0 mg/mL methanol

Cap 17, Step 2; (R)-2-(4-Hydroxy-4-phenylpiperidin-1-yl)-2-phenylaceticacid: To a solution of(S)-1-phenylethyl(R)-2-(4-hydroxy-4-phenylpiperidin-1-yl)-2-phenylacetate(0.350 g, 0.84 mmol) in dichloromethane (5 mL) was added trifluoroaceticacid (1 mL) and the mixture was stirred at room temperature for 2 hours.The volatiles were subsequently removed in vacuo and the residue waspurified by reverse-phase preparative HPLC (Primesphere C-18, 20×100 mm;CH₃CN—H₂O-0.1% TFA) to give the title compound (as TFA salt) as a whitesolid (0.230 g, 88%). LCMS: Anal. Calcd. for C₁₉H₂₁NO₃: 311.15. found:312 (M+H)⁺.

The following carboxylic acids were prepared in optically pure form in asimilar fashion:

Cap-17a

RT ₌ 2.21 (Cond'n II); ¹HNMR (500 MHz, DMSO-d₆) δ ppm 2.20-2.35 (m, 2H)2.34-2.47 (m, 2H) 3.37 (s, 4H) 3.71 (s, 1H) 5.06 (s, 2H) 7.06-7.53 (m,10H). LCMS: Anal. Calcd. for: C₂₀H₂₂N₂O₄ 354.16; Found: 355.38 (M + H)⁺.Cap-17b

RT ₌ 0.27 (Cond'n III); LCMS: Anal. Calcd. for: C₁₂H₁₄N₂O₃ 234.10;Found: 235.22 (M + H)⁺. Cap-17c

RT₌ 0.48 (Cond'n II); LCMS: Anal. Calcd. for: C₁₃H₁₈N₂O₂ 234.14; Found:235.31 (M + H)⁺. Cap-17d

RT ₌ 2.21 (Cond'n I); LCMS: Anal. Calcd. for: C₁₉H₂₁NO₂ 295.16; Found:296.33 (M + H)⁺.LCMS Conditions for Determining Retention TimeCondition IColumn: Phenomenex-Luna 4.6×50 mm S10Start % B=0Fianl % B=100Gradient Time=4 minFlow Rate=4 mL/minWavelength=220Solvent A=10% methanol-90% H₂O-0.1% TFASolvent B=90% methanol-10% H₂O-0.1% TFACondition IIColumn: Waters-Sunfire 4.6×50 mm S5Start % B=0Fianl % B=100Gradient Time=2 minFlow Rate=4 mL/minWavelength=220Solvent A=10% methanol-90% H₂O-0.1% TFASolvent B=90% methanol-10% H₂O-0.1% TFACondition IIIColumn: Phenomenex 10μ 3.0×50 mmStart % B=0Fianl % B=100Gradient Time=2 minFlow Rate=4 mL/minWavelength=220Solvent A=10% methanol-90% H₂O-0.1% TFASolvent B=90% methanol-10% H₂O-0.1% TFA

Cap-18

Step 1

(R,S)-Ethyl 2-(4-pyridyl)-2-bromoacetate: To a solution of ethyl4-pyridylacetate (1.00 g, 6.05 mmol) in dry THF (150 mL) at 0° C. underargon was added DBU (0.99 mL, 6.66 mmol). The reaction mixture wasallowed to warm to room temperature over 30 minutes and then it wascooled to −78° C. To this mixture was added CBr₄ (2.21 g, 6.66 mmol) andstirring was continued at −78° C. for 2 hours. The reaction mixture wasthen quenched with sat. aq. NH₄Cl and the phases were separated. Theorganic phase was washed (brine), dried (Na₂SO₄), filtered, andconcentrated in vacuo. The resulting yellow oil was immediately purifiedby flash chromatography (SiO₂/hexane-ethyl acetate, 1:1) to provide thetitle compound (1.40 g, 95%) as a somewhat unstable yellow oil. ¹H NMR(400 MHz, CDCl₃) δ 8.62 (dd, J=4.6, 1.8 Hz, 2H), 7.45 (dd, J=4.6, 1.8Hz, 2H), 5.24 (s, 1H), 4.21-4.29 (m, 2H), 1.28 (t, J=7.1 Hz, 3H). LCMS:Anal. Calcd. for C₉H₁₀BrNO₂: 242, 244. found: 243, 245 (M+H)⁺.

Step 2

(R,S)-Ethyl 2-(4-pyridyl)-2-(N,N-dimethylamino)acetate: To a solution of(R,S)-ethyl 2-(4-pyridyl)-2-bromoacetate (1.40 g, 8.48 mmol) in DMF (10mL) at room temperature was added dimethylamine (2M in THF, 8.5 mL, 17.0mmol). After completion of the reaction (as judged by thin layerchromatography) the volatiles were removed in vacuo and the residue waspurified by flash chromatography (Biotage, 40+M SiO₂ column; 50%-100%ethyl acetate-hexane) to provide the title compound (0.539 g, 31%) as alight yellow oil. ¹H NMR (400 MHz, CDCl₃) δ 8.58 (d, J=6.0 Hz, 2H), 7.36(d, J=6.0 Hz, 2H), 4.17 (m, 2H), 3.92 (s, 1H), 2.27 (s, 6H), 1.22 (t,J=7.0 Hz). LCMS: Anal. Calcd. for C₁₁H₁₆N₂O₂: 208. found: 209 (M+H)⁺.

Step 3

(R,S)-2-(4-Pyridyl)-2-(N,N-dimethylamino)acetic acid: To a solution of(R,S)-ethyl 2-(4-pyridyl)-2-(N,N-dimethylamino)acetate (0.200 g, 0.960mmol) in a mixture of THF-methanol-H₂O (1:1:1, 6 mL) was added powderedLiOH (0.120 g, 4.99 mmol) at room temperature. The solution was stirredfor 3 hours and then it was acidified to pH 6 using 1N HCl. The aqueousphase was washed with ethyl acetate and then it was lyophilized to givethe dihydrochloride of the title compound as a yellow solid (containingLiCl). The product was used as such in subsequent steps. ¹H NMR (400MHz, DMSO-d₆) δ 8.49 (d, J=5.7 Hz, 2H), 7.34 (d, J=5.7 Hz, 2H), 3.56 (s,1H), 2.21 (s, 6H).

The following examples were prepared in similar fashion using the methoddescribed above;

Cap-19

LCMS: Anal. Calcd. for C₉H₁₂N₂O₂: 180; found: 181 (M + H)⁺. Cap-20

LCMS: no ionization. ¹H NMR (400 MHz, CD₃OD) δ 8.55 (d, J = 4.3 Hz, 1H),7.84 (app t, J = 5.3 Hz, 1H), 7.61 (d, J = 7.8 Hz, 1H), 7.37 (app t, J =5.3 Hz, 1H), 4.35 (s, 1H), 2.60 (s, 6H). Cap-21

LCMS: Anal. Calcd. for C₉H₁₁ClN₂O₂: 214, 216; found: 215, 217 (M + H)⁺.Cap-22

LCMS: Anal. Calcd. for C₁₀H₁₂N₂O₄: 224; found: 225 (M + H)⁺. Cap-23

LCMS: Anal. Calcd. for C₁₄H₁₅NO₂: 229; found: 230 (M + H)⁺. Cap-24

LCMS: Anal. Calcd. for C₁₁H₁₂F₃NO₂: 247; found: 248 (M + H)⁺. Cap-25

LCMS: Anal. Calcd. for C₁₁H₁₂F₃NO₂: 247; found: 248 (M + H)⁺. Cap-26

LCMS: Anal. Calcd. for C₁₀H₁₂FNO₂: 197; found: 198 (M + H)⁺. Cap-27

LCMS: Anal. Calcd. for C₁₀H₁₂FNO₂: 247; found: 248 (M + H)⁺. Cap-28

LCMS: Anal. Calcd. for C₁₀H₁₂ClNO₂: 213; found: 214 (M + H)⁺. Cap-29

LCMS: Anal. Calcd. for C₁₀H₁₂ClNO₂: 213; found: 214 (M + H)⁺. Cap-30

LCMS: Anal. Calcd. for C₁₀H₁₂CINO₂: 213; found: 214 (M + H)⁺. Cap-31

LCMS: Anal. Calcd. for C₈H₁₂N₂O₂S: 200; found: 201 (M + H)⁺. Cap-32

LCMS: Anal. Calcd. for C₈H₁₁NO₂S: 185; found: 186 (M + H)⁺. Cap-33

LCMS: Anal. Calcd. for C₈H₁₁NO₂S: 185; found: 186 (M + H)⁺. Cap-34

LCMS: Anal. Calcd. for C₁₁H₁₂N₂O₃: 220; found: 221 (M + H)⁺. Cap-35

LCMS: Anal. Calcd. for C₁₂H₁₃NO₂S: 235; found: 236 (M + H)⁺. Cap-36

LCMS: Anal. Calcd. for C₁₂H₁₄N₂O₂S: 250; found: 251 (M + H)⁺.

Cap-37

Step 1

(R,S)-Ethyl 2-(quinolin-3-yl)-2-(N,N-dimethylamino)-acetate: A mixtureof ethyl N,N-dimethylaminoacetate (0.462 g, 3.54 mmol), K₃PO₄ (1.90 g,8.95 mmol), Pd(t-Bu₃P)₂ (0.090 g, 0.176 mmol) and toluene (10 mL) wasdegassed with a stream of Ar bubbles for 15 minutes. The reactionmixture was then heated at 100° C. for 12 hours, after which it wascooled to room temperature and poured into H₂O. The mixture wasextracted with ethyl acetate (2×) and the combined organic phases werewashed (H₂O, brine), dried (Na₂SO₄), filtered, and concentrated invacuo. The residue was purified first by reverse-phase preparative HPLC(Primesphere C-18, 30×100 mm; CH₃CN—H₂O-5 mM NH₄OAc) and then by flashchromatography (SiO₂/hexane-ethyl acetate, 1:1) to provide the titlecompound (0.128 g, 17%) as an orange oil. ¹H NMR (400 MHz, CDCl₃) δ 8.90(d, J=2.0 Hz, 1H), 8.32 (d, J=2.0 Hz, 1H), 8.03-8.01 (m, 2H), 7.77 (ddd,J=8.3, 6.8, 1.5 Hz, 1H), 7.62 (ddd, J=8.3, 6.8, 1.5 Hz, 1H), 4.35 (s,1H), 4.13 (m, 2H), 2.22 (s, 6H), 1.15 (t, J=7.0 Hz, 3H). LCMS: Anal.Calcd. for C₁₅H₁₈N₂O₂: 258. found: 259 (M+H)⁺.

Step 2

(R,S) 2-(Quinolin-3-yl)-2-(N,N-dimethylamino)acetic acid: A mixture of(R,S)-ethyl 2-(quinolin-3-yl)-2-(N,N-dimethylamino)acetate (0.122 g,0.472 mmol) and 6M HCl (3 mL) was heated at 100° C. for 12 hours. Thesolvent was removed in vacuo to provide the dihydrochloride of the titlecompound (0.169 g, >100%) as a light yellow foam. The unpurifiedmaterial was used in subsequent steps without further purification.LCMS: Anal. Calcd. for C₁₃H₁₄N₂O₂: 230. found: 231 (M+H)⁺.

Cap-38

Step 1

(R)—((S)-1-phenylethyl) 2-(dimethylamino)-2-(2-fluorophenyl)acetate and(S)—((S)-1-phenylethyl) 2-(dimethylamino)-2-(2-fluorophenyl)acetate: Toa mixture of (RS)-2-(dimethylamino)-2-(2-fluorophenyl)acetic acid (2.60g, 13.19 mmol), DMAP (0.209 g, 1.71 mmol) and (S)-1-phenylethanol (2.09g, 17.15 mmol) in CH₂Cl₂ (40 mL) was added EDCI (3.29 g, 17.15 mmol) andthe mixture was allowed to stir at room temperature for 12 hours. Thesolvent was then removed in vacuo and the residue partitioned with ethylacetate-H₂O. The layers were separated, the aqueous layer wasback-extracted with ethyl acetate (2×) and the combined organic phaseswere washed (H₂O, brine), dried (Na₂SO₄), filtered, and concentrated invacuo. The residue was purified by silica gel chromatography(Biotage/0-50% diethyl ether-hexane). The resulting pure diastereomericmixture was then separated by reverse-phase preparative HPLC(Primesphere C-18, 30×100 mm; CH₃CN—H₂O-0.1% TFA) to give first(S)-1-phenethyl(R)-2-(dimethylamino)-2-(2-fluorophenyl)acetate (0.501 g,13%) and then(S)-1-phenethyl(S)-2-(dimethylamino)-2-(2-fluorophenyl)-acetate (0.727g. 18%), both as their TFA salts. (S,R)-isomer: ¹H NMR (400 MHz, CD₃OD)δ 7.65-7.70 (m, 1H), 7.55-7.60 (ddd, J=9.4, 8.1, 1.5 Hz, 1H), 7.36-7.41(m, 2H), 7.28-7.34 (m, 5H), 6.04 (q, J=6.5 Hz, 1H), 5.60 (s, 1H), 2.84(s, 6H), 1.43 (d, J=6.5 Hz, 3H). LCMS: Anal. Calcd. for C₁₈H₂₀FNO₂: 301.found: 302 (M+H)⁺; (S,S)-isomer: ¹H NMR (400 MHz, CD₃OD) δ 7.58-7.63 (m,1H), 7.18-7.31 (m, 6H), 7.00 (dd, J=8.5, 1.5 Hz, 2H), 6.02 (q, J=6.5 Hz,1H), 5.60 (s, 1H), 2.88 (s, 6H), 1.54 (d, J=6.5 Hz, 3H). LCMS: Anal.Calcd. for C₁₈H₂₀FNO₂: 301. found: 302 (M+H)⁺.

Step 2

(R)-2-(dimethylamino)-2-(2-fluorophenyl)acetic acid: A mixture of(R)—((S)-1-phenylethyl) 2-(dimethylamino)-2-(2-fluorophenyl)acetate TFAsalt (1.25 g, 3.01 mmol) and 20% Pd(OH)₂/C (0.125 g) in ethanol (30 mL)was hydrogenated at room temperature and atmospheric pressure (H₂balloon) for 4 hours. The solution was then purged with Ar, filteredthrough diatomaceous earth (Celite®), and concentrated in vacuo. Thisgave the title compound as a colorless solid (0.503 g, 98%). ¹H NMR (400MHz, CD₃OD) δ 7.53-7.63 (m, 2H), 7.33-7.38 (m, 2H), 5.36 (s, 1H), 2.86(s, 6H). LCMS: Anal. Calcd. for C₁₀H₁₂FNO₂: 197. found: 198 (M+H)⁺.

The S-isomer could be obtained from (S)—((S)-1-phenylethyl)2-(dimethylamino)-2-(2-fluorophenyl)acetate TFA salt in similar fashion.

Cap-39

A mixture of (R)-(2-chlorophenyl)glycine (0.300 g, 1.62 mmol),formaldehyde (35% aqueous solution, 0.80 mL, 3.23 mmol) and 20%Pd(OH)₂/C (0.050 g) was hydrogenated at room temperature and atmosphericpressure (H₂ balloon) for 4 hours. The solution was then purged with Ar,filtered through diatomaceous earth (Celite®) and concentrated in vacuo.The residue was purified by reverse-phase preparative HPLC (PrimesphereC-18, 30×100 mm; CH₃CN—H₂O-0.1% TFA) to give theTFA salt of the titlecompound (R)-2-(dimethylamino)-2-(2-chlorophenyl)acetic acid as acolorless oil (0.290 g, 55%). ¹H NMR (400 MHz, CD₃OD) δ 7.59-7.65 (m,2H), 7.45-7.53 (m, 2H), 5.40 (s, 1H), 2.87 (s, 6H). LCMS: Anal. Calcd.for C₁₀H₁₂ClNO₂: 213. found: 214 (M+H)⁺.

Cap-40

To an ice-cold solution of (R)-(2-chlorophenyl)glycine (1.00 g, 5.38mmol) and NaOH (0.862 g, 21.6 mmol) in H₂O (5.5 mL) was added methylchloroformate (1.00 mL, 13.5 mmol) dropwise. The mixture was allowed tostir at 0° C. for 1 hour and then it was acidified by the addition ofconc. HCl (2.5 mL). The mixture was extracted with ethyl acetate (2×)and the combined organic phase was washed (H₂O, brine), dried (Na₂SO₄),filtered, and concentrated in vacuo to give the title compound(R)-2-(methoxycarbonylamino)-2-(2-chlorophenyl)acetic acid as ayellow-orange foam (1.31 g, 96%). ¹H NMR (400 MHz, CD₃OD) δ 7.39-7.43(m, 2H), 7.29-7.31 (m, 2H), 5.69 (s, 1H), 3.65 (s, 3H). LCMS: Anal.Calcd. for C₁₀H₁₀ClNO₄: 243. found: 244 (M+H)⁺.

Cap-41

To a suspension of 2-(2-(chloromethyl)phenyl)acetic acid (2.00 g, 10.8mmol) in THF (20 mL) was added morpholine (1.89 g, 21.7 mmol) and thesolution was stirred at room temperature for 3 hours. The reactionmixture was then diluted with ethyl acetate and extracted with H₂O (2×).The aqueous phase was lyophilized and the residue was purified by silicagel chromatography (Biotage/0-10% methanol-CH₂Cl₂) to give the titlecompound 2-(2-(Morpholinomethyl)phenyl)acetic acid as a colorless solid(2.22 g, 87%). ¹H NMR (400 MHz, CD₃OD) δ 7.37-7.44 (m, 3H), 7.29-7.33(m, 1H), 4.24 (s, 2H), 3.83 (br s, 4H), 3.68 (s, 2H), 3.14 (br s, 4H).LCMS: Anal. Calcd. for C₁₃H₁₇NO₃: 235. found: 236 (M+H)⁺.

The following examples were similarly prepared using the methoddescribed for Cap-41:

Cap-42

LCMS: Anal. Calcd. for C₁₄H₁₉NO₂: 233; found: 234 (M + H)⁺. Cap-43

LCMS: Anal. Calcd. for C₁₃H₁₇NO₂: 219; found: 220 (M + H)⁺. Cap-44

LCMS: Anal. Calcd. for C₁₁H₁₅NO₂: 193; found: 194 (M + H)⁺. Cap-45

LCMS: Anal. Calcd. for C₁₄H₂₀N₂O₂: 248; found: 249 (M + H)⁺.

Cap-45a

HMDS (1.85 mL, 8.77 mmol) was added to a suspension of(R)-2-amino-2-phenylacetic acid p-toluenesulfonate (2.83 g, 8.77 mmol)in CH₂Cl₂ (10 mL) and the mixture was stirred at room temperature for 30minutes. Methyl isocyanate (0.5 g, 8.77 mmol) was added in one portionstirring continued for 30 minutes. The reaction was quenched by additionof H₂O (5 mL) and the resulting precipitate was filtered, washed withH₂O and n-hexanes, and dried under vacuum.(R)-2-(3-methylureido)-2-phenylacetic acid (1.5 g; 82%) was recovered asa white solid and it was used without further purification. ¹H NMR (500MHz, DMSO-d₆) δ ppm 2.54 (d, J=4.88 Hz, 3H) 5.17 (d, J=7.93 Hz, 1H) 5.95(q, J=4.48 Hz, 1H) 6.66 (d, J=7.93 Hz, 1H) 7.26-7.38 (m, 5H) 12.67 (s,1H). LCMS: Anal. Calcd. for C₁₀H₁₂N₂O₃ 208.08. found 209.121 (M+H)⁺.HPLC Phenomenex C-18 3.0×46 mm, 0 to 100% B over 2 minutes, 1 minutehold time, A=90% water, 10% methanol, 0.1% TFA, B=10% water, 90%methanol, 0.1% TFA, RT=1.38 min, 90% homogeneity index.

Cap-46

The desired product was prepared according to the method described forCap-45a. ¹H NMR (500 MHz, DMSO-d₆) δ ppm 0.96 (t, J=7.17 Hz, 3H)2.94-3.05 (m, 2H) 5.17 (d, J=7.93 Hz, 1H) 6.05 (t, J=5.19 Hz, 1H) 6.60(d, J=7.63 Hz, 1H) 7.26-7.38 (m, 5H) 12.68 (s, 1H). LCMS: Anal. Calcd.for C₁₁H₁₄N₂O₃ 222.10 found 223.15 (M+H)⁺. HPLC XTERRA C-18 3.0×506 mm,0 to 100% B over 2 minutes, 1 minute hold time, A=90% water, 10%methanol, 0.2% H₃PO₄, B=10% water, 90% methanol, 0.2% H₃PO₄, RT=0.87min, 90% homogeneity index.

Cap-47

Step 1

(R)-tert-butyl 2-(3,3-dimethylureido)-2-phenylacetate: To a stirredsolution of (R)-tert-butyl-2-amino-2-phenylacetate (1.0 g, 4.10 mmol)and Hunig's base (1.79 mL, 10.25 mmol) in DMF (40 mL) was addeddimethylcarbamoyl chloride (0.38 mL, 4.18 mmol) dropwise over 10minutes. After stirring at room temperature for 3 hours, the reactionwas concentrated under reduced pressure and the resulting residue wasdissolved in ethyl acetate. The organic layer was washed with H₂O, 1Naq. HCl and brine, dried (MgSO₄), filtered and concentrated underreduced pressure. (R)-tert-butyl 2-(3,3-dimethylureido)-2-phenylacetatewas obtained as a white solid (0.86 g; 75%) and used without furtherpurification. ¹H NMR (500 MHz, DMSO-d₆) δ ppm 1.33 (s, 9H) 2.82 (s, 6H)5.17 (d, J=7.63 Hz, 1H) 6.55 (d, J=7.32 Hz, 1H) 7.24-7.41 (m, 5H). LCMS:Anal. Calcd. for C₁₅H₂₂N₂O₃ 278.16 found 279.23 (M+H)⁺; HPLC PhenomenexLUNA C-18 4.6×50 mm, 0 to 100% B over 4 minutes, 1 minute hold time,A=90% water, 10% methanol, 0.1% TFA, B=10% water, 90% methanol, 0.1%TFA, RT=2.26 min, 97% homogeneity index.

Step 2

(R)-2-(3,3-dimethylureido)-2-phenylacetic acid: To a stirred solution of((R)-tert-butyl 2-(3,3-dimethylureido)-2-phenylacetate (0.86 g, 3.10mmol) in CH₂Cl₂ (250 mL) was added TFA (15 mL) dropwise and theresulting solution was stirred at rt for 3 hours. The desired compoundwas then precipitated out of solution with a mixture of EtOAC:Hexanes(5:20), filtered off and dried under reduced pressure.(R)-2-(3,3-dimethylureido)-2-phenylacetic acid was isolated as a whitesolid (0.59 g, 86%) and used without further purification. ¹H NMR (500MHz, DMSO-d₆) δ ppm 2.82 (s, 6H) 5.22 (d, J=7.32 Hz, 1H) 6.58 (d, J=7.32Hz, 1H) 7.28 (t, J=7.17 Hz, 1H) 7.33 (t, J=7.32 Hz, 2H) 7.38-7.43 (m,2H) 12.65 (s, 1H). LCMS: Anal. Calcd. for C₁₁H₁₄N₂O₃: 222.24. found:223.21 (M+H)⁺. HPLC XTERRA C-18 3.0×50 mm, 0 to 100% B over 2 minutes, 1minute hold time, A=90% water, 10% methanol, 0.2% H₃PO₄, B=10% water,90% methanol, 0.2% H₃PO₄, RT=0.75 min, 93% homogeneity index.

Cap-48

Step 1

(R)-tert-butyl 2-(3-cyclopentylureido)-2-phenylacetate: To a stirredsolution of (R)-2-amino-2-phenylacetic acid hydrochloride (1.0 g, 4.10mmol) and Hunig's base (1.0 mL, 6.15 mmol) in DMF (15 mL) was addedcyclopentyl isocyanate (0.46 mL, 4.10 mmol) dropwise and over 10minutes. After stirring at room temperature for 3 hours, the reactionwas concentrated under reduced pressure and the resulting residue wastraken up in ethyl acetate. The organic layer was washed with H₂O andbrine, dried (MgSO₄), filtered, and concentrated under reduced pressure.(R)-tert-butyl 2-(3-cyclopentylureido)-2-phenylacetate was obtained asan opaque oil (1.32 g; 100%) and used without further purification. ¹HNMR (500 MHz, CD₃Cl-D) δ ppm 1.50-1.57 (m, 2H) 1.58-1.66 (m, 2H)1.87-1.97 (m, 2H) 3.89-3.98 (m, 1H) 5.37 (s, 1H) 7.26-7.38 (m, 5H).LCMS: Anal. Calcd. for C₁₈H₂₆N₂O₃ 318.19 found 319.21 (M+H)⁺; HPLCXTERRA C-18 3.0×50 mm, 0 to 100% B over 4 minutes, 1 minute hold time,A=90% water, 10% methanol, 0.1% TFA, B=10% water, 90% methanol, 0.1%TFA, RT=2.82 min, 96% homogeneity index.

Step 2

(R)-2-(3-cyclopentylureido)-2-phenylacetic acid: To a stirred solutionof (R)-tert-butyl 2-(3-cyclopentylureido)-2-phenylacetate (1.31 g, 4.10mmol) in CH₂Cl₂ (25 mL) was added TFA (4 mL) and trietheylsilane (1.64mL; 10.3 mmol) dropwise, and the resulting solution was stirred at roomtemperature for 6 hours. The volatile components were removed underreduced pressure and the crude product was recrystallized in ethylacetate/pentanes to yield (R)-2-(3-cyclopentylureido)-2-phenylaceticacid as a white solid (0.69 g, 64%). ¹H NMR (500 MHz, DMSO-d₆) δ ppm1.17-1.35 (m, 2H) 1.42-1.52 (m, 2H) 1.53-1.64 (m, 2H) 1.67-1.80 (m, 2H)3.75-3.89 (m, 1H) 5.17 (d, J=7.93 Hz, 1H) 6.12 (d, J=7.32 Hz, 1H) 6.48(d, J=7.93 Hz, 1H) 7.24-7.40 (m, 5H) 12.73 (s, 1H). LCMS: Anal. Calcd.for C₁₄H₁₈N₂O₃: 262.31. found: 263.15 (M+H)⁺. HPLC XTERRA C-18 3.0×50mm, 0 to 100% B over 2 minutes, 1 minute hold time, A=90% water, 10%methanol, 0.2% H₃PO₄, B=10% water, 90% methanol, 0.2% H₃PO₄, RT=1.24min, 100% homogeneity index.

Cap-49

To a stirred solution of 2-(benzylamino)acetic acid (2.0 g, 12.1 mmol)in formic acid (91 mL) was added formaldehyde (6.94 mL, 93.2 mmol).After five hours at 70° C., the reaction mixture was concentrated underreduced pressure to 20 mL and a white solid precipitated. Followingfiltration, the mother liquors were collected and further concentratedunder reduced pressure providing the crude product. Purification byreverse-phase preparative HPLC (Xterra 30×100 mm, detection at 220 nm,flow rate 35 mL/min, 0 to 35% B over 8 min; A=90% water, 10% methanol,0.1% TFA, B=10% water, 90% methanol, 0.1% TFA) provided the titlecompound 2-(benzyl(methyl)-amino)acetic acid as its TFA salt (723 mg,33%) as a colorless wax. ¹H NMR (300 MHz, DMSO-d₆) δ ppm 2.75 (s, 3H)4.04 (s, 2H) 4.34 (s, 2H) 7.29-7.68 (m, 5H). LCMS: Anal. Calcd. for:C₁₀H₁₃NO₂ 179.09. Found: 180.20 (M+H)⁺.

Cap-50

To a stirred solution of 3-methyl-2-(methylamino)butanoic acid (0.50 g,3.81 mmol) in water (30 mL) was added K₂CO₃ (2.63 g, 19.1 mmol) andbenzyl chloride (1.32 g, 11.4 mmol). The reaction mixture was stirred atambient temperature for 18 hours. The reaction mixture was extractedwith ethyl acetate (30 mL×2) and the aqueous layer was concentratedunder reduced pressure providing the crude product which was purified byreverse-phase preparative HPLC (Xterra 30×100 mm, detection at 220 nm,flow rate 40 mL/min, 20 to 80% B over 6 min; A=90% water, 10% methanol,0.1% TFA, B=10% water, 90% methanol, 0.1% TFA) to provide2-(benzyl(methyl)amino)-3-methylbutanoic acid, TFA salt (126 mg, 19%) asa colorless wax. ¹H NMR (500 MHz, DMSO-d₆) δ ppm 0.98 (d, 3H) 1.07 (d,3H) 2.33-2.48 (m, 1H) 2.54-2.78 (m, 3H) 3.69 (s, 1H) 4.24 (s, 2H)7.29-7.65 (m, 5H). LCMS: Anal. Calcd. for: C₁₃H₁₉NO₂ 221.14. Found:222.28 (M+H)⁺.

Cap-51

Na₂CO₃ (1.83 g, 17.2 mmol) was added to NaOH (33 mL of 1M/H₂O, 33 mmol)solution of L-valine (3.9 g, 33.29 mmol) and the resulting solution wascooled with ice-water bath. Methyl chloroformate (2.8 mL, 36.1 mmol) wasadded dropwise over 15 min, the cooling bath was removed and thereaction mixture was stirred at ambient temperature for 3.25 hr. Thereaction mixture was washed with ether (50 mL, 3×), and the aqueousphase was cooled with ice-water bath and acidified with concentrated HClto a pH region of 1-2, and extracted with CH₂Cl₂ (50 mL, 3×). Theorganic phase was dried (MgSO₄) and evaporated in vacuo to afford Cap-51as a white solid (6 g). ¹H NMR for the dominant rotamer (DMSO-d₆, δ=2.5ppm, 500 MHz): 12.54 (s, 1H), 7.33 (d, J=8.6, 1H), 3.84 (dd, J=8.4, 6.0,1H), 3.54 (s, 3H), 2.03 (m, 1H), 0.87 (m, 6H). HRMS: Anal. Calcd. for[M+H]⁺ C₂H₁₄NO₄: 176.0923. found 176.0922.

Cap 51 (Alternate Route)

DIEA (137.5 mL, 0.766 mol) was added to a suspension of (S)-tert-butyl2-amino-3-methylbutanoate hydrochloride (75.0 g, 0.357 mol) in THF (900mL), and the mixture was cooled to 0° C. (ice/water bath). Methylchloroformate (29.0 mL, 0.375 mol) was added dropwise over 45 min, thecooling bath was removed and the heterogeneous mixture was stirred atambient temperature for 3 h. The solvent was removed under diminishedpressure and the residue partitioned between EtOAc and water (1 L each).The organic layer was washed with H₂O (1 L) and brine (1 L), dried(MgSO₄), filtered and concentrated under diminished pressure. The crudematerial was passed through a plug of silica gel (1 kg), eluting withhexanes (4 L) and 15:85 EtOAc/hexanes (4 L) to afford (S)-tert-butyl2-(methoxycarbonylamino)-3-methylbutanoate as a clear oil (82.0 g, 99%yield). ¹H-NMR (500 MHz, DMSO-d₆, δ=2.5 ppm) 7.34 (d, J=8.6, 1H), 3.77(dd, J=8.6, 6.1, 1H), 3.53 (s, 3H), 1.94-2.05 (m, 1H), 1.39 (s, 9H),0.83-0.92 (m, 6H). ¹³C-NMR (126 MHz, DMSO-d₆, δ=39.2 ppm) 170.92,156.84, 80.38, 60.00, 51.34, 29.76, 27.62, 18.92, 17.95. LC/MS: [M+Na]⁺254.17.

Trifluoroacetic acid (343 mL, 4.62 mol) and Et₃SiH (142 mL, 0.887 mol)were added sequentially to a solution of (S)-tert-butyl2-(methoxycarbonylamino)-3-methylbutanoate (82.0 g, 0.355 mol) in CH₂Cl₂(675 mL), and the mixture was stirred at ambient temperature for 4 h.The volatile component was removed under diminished pressure and theresultant oil triturated with petroleum ether (600 mL) to afford a whitesolid, which was filtered and washed with hexanes (500 mL) and petroleumether (500 mL). Recrystallization from EtOAc/petroleum ether affordedCap-51 as white flaky crystals (54.8 g, 88% yield). MP=108.5-109.5° C.¹H NMR (500 MHz, DMSO-d₆, δ=2.5 ppm) 12.52 (s, 1H), 7.31 (d, J=8.6, 1H),3.83 (dd, J=8.6, 6.1, 1H), 3.53 (s, 3 H), 1.94-2.07 (m, 1H), 0.86 (dd,J=8.9, 7.0, 6H). ¹³C NMR (126 MHz, DMSO-d₆, δ=39.2 ppm) 173.30, 156.94,59.48, 51.37, 29.52, 19.15, 17.98. LC/MS: [M+H]⁺=176.11. Anal. Calcd.for C₇H₁₃NO₄: C, 47.99; H, 7.48; N, 7.99. Found: C, 48.17; H, 7.55; N,7.99. Optical Rotation: [α]_(D)=−4.16 (12.02 mg/mL; MeOH). Opticalpurity: >99.5% ee. Note: the optical purity assessment was made on themethyl ester derivative of Cap-51, which was prepared under a standardTMSCHN₂ (benzene/MeOH) esterification protocol. HPLC analyticalconditions: column, ChiralPak AD-H (4.6×250 mm, 5 μm); solvent, 95%heptane/5% IPA (isocratic); flow rate, 1 mL/min; temperature, 35° C.; UVmonitored at 205 nm.

[Note: Cap 51 could also be purchased from Flamm.]

Cap-52 (Same as Cap-12)

Cap-52 was synthesized from L-alanine according to the proceduredescribed for the synthesis of Cap-51. For characterization purposes, aportion of the crude material was purified by a reverse phaseHPLC(H₂O/methanol/TFA) to afford Cap-52 as a colorless viscous oil. ¹HNMR (DMSO-d₆, δ=2.5 ppm, 500 MHz): 12.49 (br s, 1H), 7.43 (d, J=7.3,0.88H), 7.09 (app br s, 0.12H), 3.97 (m, 1H), 3.53 (s, 3H), 1.25 (d,J=7.3, 3H).

Cap-53 to -64 were prepared from appropriate starting materialsaccording to the procedure described for the synthesis of Cap-51, withnoted modifications if any.

Cap Structure Data Cap-53a: (R) Cap-53b: (S)

¹H NMR (DMSO-d₆, δ = 2.5 ppm, 500 MHz): δ 12.51 (br s, 1H), 7.4 (d, J =7.9, 0.9H), 7.06 (app s, 0.1H), 3.86-3.82 (m, 1H), 3.53 (s,3H),1.75-1.67 (m, 1H), 1.62- 1.54 (m, 1H), 0.88 (d, J = 7.3, 3H). RT = 0.77minutes (Cond. 2); LC/MS: Anal. Calcd. for [M + Na]⁺ C₆H₁₁NNaO₄: 184.06;found 184.07. HRMS Calcd. for [M + Na]⁺ C₆H₁₁NNaO₄: 184.0586; found184.0592. Cap-54a: (R) Cap-54b: (S)

¹H NMR (DMSO-d₆, δ = 2.5 ppm. 500 MHz): 8 12.48 (s, 1H), 7.58 (d, J =7.6, 0.9H), 7.25 (app s, 0.1H), 3.52 (s, 3H), 3.36-3.33 (m, 1H),1.10-1.01 (m, 1H), 0.54-0.49 (m, 1H), 0.46-0.40 (m, 1H), 0.39-0.35 (m,1H), 0.31-0.21 (m, 1H). HRMS Calcd. for [M + H]⁺ C₇H₁₂NO₄: 174.0766;found 174.0771 Cap-55:

¹H NMR (DMSO-d₆, δ = 2.5 ppm, 500 MHz): δ 12.62 (s, 1H), 7.42 (d, J =8.2, 0.9H), 7.07 (app s, 0.1H), 5.80-5.72 (m, 1H), 5.10 (d, J = 17.1,1H), 5.04 (d, J = 10.4, 1H), 4.01-3.96 (m, 1H), 3.53 (s, 3H), 2.47-2.42(m, 1H), 2.35-2.29 (m, 1H). Cap-56:

¹H NMR (DMSO-d₆, δ = 2.5 ppm, 500 MHz): δ 12.75 (s, IH), 7.38 (d, J =8.3, 0.9H), 6.96 (app s, 0.1H), 4.20-4.16 (m, 1H), 3.60-3.55 (m, 2H),3.54 (s, 3H), 3.24 (s, 3H). Cap-57:

¹H NMR (DMSO-d₆, δ = 2.5 ppm, 500 MHz): δ 12.50 (s, 1H), 8.02 (d, J =7.7, 0.08H), 7.40 (d, J = 7.9, 0.76H), 7.19 (d, J = 8.2, 0.07H), 7.07(d, J = 6.7, 0.09H), 4.21-4.12 (m, 0.08H), 4.06-3.97 (m, 0.07H),3.96-3.80 (m, 0.85H), 3.53 (s, 3H), 1.69-1.51 (m, 2H), 1.39-1.26 (m,2H), 0.85 (t, J = 7.4, 3H). LC (Cond. 2): RT = 1.39 LC/MS: Anal. Calcd.for [M + H]⁺ C₇H₁₄NO₄: 176.09; found 176.06. Cap-58:

¹H NMR (DMSO-d₆, δ = 2.5 ppm, 500 MHz): δ 12.63 (brs, 1H), 7.35 (s, 1H),7.31 (d, J = 8.2, 1H), 6.92 (s, 1H), 4.33-4.29 (m, 1H), 3.54 (s,3H),2.54 (dd, J = 15.5, 5.4, 1H), 2.43 (dd, J = 15.6, 8.0, 1H). RT = 0.16min (Cond. 2); LC/MS: Anal. Calcd. for [M + H]⁺ C₆H₁₁N₂O₅: 191.07; found191.14. Cap-59a: (R) Cap-59b: (S)

¹H NMR (DMSO-d₆, δ = 2.5 ppm, 400 MHz): δ 12.49 (br s, 1H), 7.40 (d, J =7.3, 0.89H), 7.04 (br s, 0.11H), 4.00-3.95 (m, 3H), 1.24 (d, J = 7.3,3H), 1.15 (t, J = 7.2, 3H). HRMS: Anal. Calcd. for [M + H]⁺ C₆H₁₂NO₄:162.0766; found 162.0771. Cap-60:

The crude material was purified with a reverse phase HPLC (H₂O/MeOH/TFA)to afford a colorless viscous oil that crystallized to a white solidupon exposure to high vacuum. ¹H NMR (DMSO-d₆, δ = 2.5 ppm, 400 MHz): δ12.38 (br s, 1H), 7.74 (s, 0.82H), 7.48 (s, 0.18H), 3.54/3.51 (two s,3H), 1.30 (m, 2H), 0.98 (m, 2H). HRMS: Anal. Calcd. for [M + H]⁺C₆H₁₀NO₄: 160.0610; found 160.0604. Cap-61:

¹H NMR (DMSO-d₆, δ = 2.5 ppm, 400 MHz): δ 12.27 (brs, 1H), 7.40 (brs,1H), 3.50 (s, 3H), 1.32 (s, 6H). HRMS: Anal. Calcd. for [M + H]⁺C₆H₁₂NO₄: 162.0766; found 162.0765. Cap-62:

¹H NMR (DMSO-d₆, δ = 2.5 ppm, 400 MHz): δ 12.74 (br s, 1H), 4.21 (d, J =10.3, 0.6H), 4.05 (d, J = 10.0, 0.4H), 3.62/3.60 (two singlets, 3H), 3.0(s, 3H), 2.14-2.05 (m, 1H), 0.95 (d, J = 6.3, 3H), 0.81 (d, J = 6.6,3H). LC/MS: Anal. Calcd. for [M − H]⁻ C₈H₁₄NO₄: 188.09; found 188.05.Cap-63:

[Note: the reaction was allowed to run for longer than what was notedfor the general procedure.] ¹H NMR (DMSO-d₆, δ = 2.5 ppm, 400 MHz):12.21 (brs, 1H), 7.42 (br s, 1H), 3.50 (s, 3H), 2.02-1.85 (m, 4H),1.66-1.58 (m,4H). LC/MS: Anal. Calcd. for [M + H]⁺ C₈H₁₄NO₄: 188.09;found 188.19. Cap-64:

[Note: the reaction was allowed to run for longer than what was notedfor the general procedure.] ¹H NMR (DMSO-d₆, δ = 2.5 ppm, 400 MHz):12.35 (br s, 1H), 7.77 (s, 0.82H), 7.56/7.52 (overlapping br s, 0.18H),3.50 (s, 3H), 2.47-2.40 (m, 2H), 2.14-2.07 (m, 2H), 1.93-1.82 (m, 2H).

Cap-65

Methyl chloroformate (0.65 mL, 8.39 mmol) was added dropwise over 5 minto a cooled (ice-water) mixture of Na₂CO₃ (0.449 g, 4.23 mmol), NaOH(8.2 mL of 1M/H₂O, 8.2 mmol) and (S)-2-amino-3-hydroxy-3-methylbutanoicacid (1.04 g, 7.81 mmol). The reaction mixture was stirred for 45 min,and then the cooling bath was removed and stirring was continued for anadditional 3.75 hr. The reaction mixture was washed with CH₂Cl₂, and theaqueous phase was cooled with ice-water bath and acidified withconcentrated HCl to a pH region of 1-2. The volatile component wasremoved in vacuo and the residue was taken up in a 2:1 mixture ofMeOH/CH₂Cl₂ (15 mL) and filtered, and the filterate was rotervaped toafford Cap-65 as a white semi-viscous foam (1.236 g). ¹H NMR (DMSO-d₆,δ=2.5 ppm, 400 MHz): δ 6.94 (d, J=8.5, 0.9H), 6.53 (br s, 0.1H), 3.89(d, J=8.8, 1H), 2.94 (s, 3H), 1.15 (s, 3H), 1.13 (s, 3H).

Cap-66 and -67 were prepared from appropriate commercially availablestarting materials by employing the procedure described for thesynthesis of Cap-65.

Cap-66

¹H NMR (DMSO-d₆, δ=2.5 ppm, 400 MHz): δ 12.58 (br s, 1H), 7.07 (d,J=8.3, 0.13H), 6.81 (d, J=8.8, 0.67H), 4.10-4.02 (m, 1.15H), 3.91 (dd,J=9.1, 3.5, 0.85H), 3.56 (s, 3H), 1.09 (d, J=6.2, 3H). [Note: only thedominant signals of NH were noted].

Cap-67

¹H NMR (DMSO-d₆, δ=2.5 ppm, 400 MHz): 12.51 (br s, 1H), 7.25 (d, J=8.4,0.75H), 7.12 (br d, J=0.4, 0.05H), 6.86 (br s, 0.08H), 3.95-3.85 (m,2H), 3.54 (s, 3H), 1.08 (d, J=6.3, 3H). [Note: only the dominant signalsof NH were noted].

Cap-68

Methyl chloroformate (0.38 ml, 4.9 mmol) was added drop-wise to amixture of 1N NaOH (aq) (9.0 ml, 9.0 mmol), 1M NaHCO₃ (aq) (9.0 ml, 9.0mol), L-aspartic acid β-benzyl ester (1.0 g, 4.5 mmol) and Dioxane (9ml). The reaction mixture was stirred at ambient conditions for 3 hr,and then washed with Ethyl acetate (50 ml, 3×). The aqueous layer wasacidified with 12N HCl to a pH ˜1-2, and extracted with ethyl acetate(3×50 ml). The combined organic layers were washed with brine, dried(Na₂SO₄), filtered, and concentrated in vacuo to afford Cap-68 as alight yellow oil (1.37 g; mass is above theoretical yield, and theproduct was used without further purification). ¹H NMR (DMSO-d₆, δ=2.5ppm, 500 MHz): δ 12.88 (br s, 1H), 7.55 (d, J=8.5, 1H), 7.40-7.32 (m,5H), 5.13 (d, J=12.8, 1H), 5.10 (d, J=12.9, 1H), 4.42-4.38 (m, 1H), 3.55(s, 3H), 2.87 (dd, J=16.2, 5.5, 1H), 2.71 (dd, J=16.2, 8.3, 1H). LC(Cond. 2): RT=1.90 min; LC/MS: Anal. Calcd. For [M+H]⁺ C₁₃H₁₆NO₆:282.10. found 282.12.

Cap-69a and -69b

NaCNBH₃ (2.416 g, 36.5 mmol) was added in batches to a chilled (˜15° C.)water (17 mL)/MeOH (10 mL) solution of alanine (1.338 g, 15.0 mmol). Afew minutes later acetaldehyde (4.0 mL, 71.3 mmol) was added drop-wiseover 4 min, the cooling bath was removed, and the reaction mixture wasstirred at ambient condition for 6 hr. An additional acetaldehyde (4.0mL) was added and the reaction was stirred for 2 hr. Concentrated HClwas added slowly to the reaction mixture until the pH reached ˜1.5, andthe resulting mixture was heated for 1 hr at 40° C. Most of the volatilecomponent was removed in vacuo and the residue was purified with aDowex® 50WX8-100 ion-exchange resin (column was washed with water, andthe compound was eluted with dilute NH₄OH, prepared by mixing 18 ml ofNH₄OH and 282 ml of water) to afford Cap-69 (2.0 g) as an off-white softhygroscopic solid. ¹H NMR (DMSO-d₆, δ=2.5 ppm, 400 MHz): δ 3.44 (q,J=7.1, 1H), 2.99-2.90 (m, 2H), 2.89-2.80 (m, 2H), 1.23 (d, J=7.1, 3H),1.13 (t, J=7.3, 6H).

Cap-70 to -74x were prepared according to the procedure described forthe synthesis of Cap-69 by employing appropriate starting materials.

Cap-70a: (R) Cap-70b: (S)

1H NMR (DMSO-d₆, δ = 2.5 ppm, 400 MHz): δ 3.42 (q, J = 7.1, 1H),2.68-2.60 (m, 4H), 1.53-1.44 (m, 4H), 1.19 (d, J = 7.3, 3H), 0.85 (t, J= 7.5, 6H). LC/MS: Anal. Calcd. for [M + H]⁺ C₉H₂₀NO₂: 174.15; found174.13. Cap-71a: (R) Cap-71b: (S)

¹H NMR (DMSO-d₆, δ = 2.5 ppm, 500 MHz): δ 3.18-3.14 (m, 1H), 2.84-2.77(m, 2H), 2.76-2.68 (m, 2H), 1.69-1.54 (m, 2H), 1.05 (t, J = 7.2, 6H),0.91 (t, J = 7.3, 3H). LC/MS: Anal. Calcd. for [M + H]⁺ C₈H₁₈NO₂:160.13; found 160.06. Cap-72:

¹H NMR (DMSO-d₆, δ = 2.5 ppm, 400 MHz): δ 2.77-2.66 (m, 3H), 2.39-2.31(m, 2H), 1.94-1.85 (m, 1H), 0.98 (t, J = 7.1, 6H), 0.91 (d, J = 6.5,3H), 0.85 (d, J = 6.5, 3H). LC/MS: Anal. Calcd. for [M + H]⁺ C₉H₂₀NO₂:174.15; found 174.15. Cap-73:

¹H NMR (DMSO-d₆, δ = 2.5 ppm, 500 MHz): δ 9.5 (br s, 1H). 3.77 (dd, J =10.8, 4.1, 1H), 3.69-3.61 ( m, 2H), 3.26 (s, 3H), 2.99-2.88 (m, 4H),1.13 (t, J = 7.2, 6H). Cap-74:

¹H NMR (DMSO-d₆, δ = 2.5 ppm, 500 MHz): δ 7.54 (s, 1H), 6.89 (s, 1H),3.81 (t, J = 6.6, k, 1H), 2.82-2.71 (m, 4H), 2.63 (dd, J = 15.6, 7.0,1H), 2.36 (dd, J = 15.4,6.3, 1H), 1.09 (t, J = 7.2, 6H). RT = 0.125minutes (Cond. 2); LC/MS: Anal. Calcd. for [M + H]⁺ C₈H₁₇N₂O₃: 189.12;found 189.13. Cap-74x:

LC/MS: Anal. Calcd. for [M + H]⁺ C₁₀H₂₂NO₂: 188.17; found 188.21

Cap-75

Cap-75, step a

NaBH₃CN (1.6 g, 25.5 mmol) was added to a cooled (ice/water bath) water(25 ml)/methanol (15 ml) solution of H-D-Ser-OBz1 HCl (2.0 g, 8.6 mmol).Acetaldehyde (1.5 ml, 12.5 mmol) was added drop-wise over 5 min, thecooling bath was removed, and the reaction mixture was stirred atambient condition for 2 hr. The reaction was carefully quenched with 12NHCl and concentrated in vacuo. The residue was dissolved in water andpurified with a reverse phase HPLC (MeOH/H₂O/TFA) to afford the TFA saltof (R)-benzyl 2-(diethylamino)-3-hydroxypropanoate as a colorlessviscous oil (1.9 g). ¹H NMR (DMSO-d₆, δ=2.5 ppm, 500 MHz): δ 9.73 (br s,1H), 7.52-7.36 (m, 5H), 5.32 (d, J=12.2, 1H), 5.27 (d, J=12.5, 1H),4.54-4.32 (m, 1H), 4.05-3.97 (m, 2H), 3.43-3.21 (m, 4H), 1.23 (t, J=7.2,6H). LC/MS (Cond. 2): RT=1.38 min; LC/MS: Anal. Calcd. for [M+H]⁺C₁₄H₂₂NO₃: 252.16. found 252.19.

Cap-75

NaH (0.0727 g, 1.82 mmol, 60%) was added to a cooled (ice-water) THF(3.0 mL) solution of the TFA salt (R)-benzyl2-(diethylamino)-3-hydroxypropanoate (0.3019 g, 0.8264 mmol) preparedabove, and the mixture was stirred for 15 min. Methyl iodide (56 μL,0.90 mmol) was added and stirring was continued for 18 hr while allowingthe bath to thaw to ambient condition. The reaction was quenched withwater and loaded onto a MeOH pre-conditioned MCX (6 g) cartridge, andwashed with methanol followed by compound elution with 2N NH₃/Methanol.Removal of the volatile component in vacuo afforded Cap-75, contaminatedwith (R)-2-(diethylamino)-3-hydroxypropanoic acid, as a yellowsemi-solid (100 mg). The product was used as is without furtherpurification.

Cap-76

NaCNBH₃ (1.60 g, 24.2 mmol) was added in batches to a chilled (˜15° C.)water/MeOH (12 mL each) solution of(S)-4-amino-2-(tert-butoxycarbonylamino) butanoic acid (2.17 g, 9.94mmol). A few minutes later acetaldehyde (2.7 mL, 48.1 mmol) was addeddrop-wise over 2 min, the cooling bath was removed, and the reactionmixture was stirred at ambient condition for 3.5 hr. An additionalacetaldehyde (2.7 mL, 48.1 mmol) was added and the reaction was stirredfor 20.5 hr. Most of the MeOH component was removed in vacuo, and theremaining mixture was treated with concentrated HCl until its pH reached˜1.0 and then heated for 2 hr at 40° C. The volatile component wasremoved in vacuo, and the residue was treated with 4 M HCl/dioxane (20mL) and stirred at ambient condition for 7.5 hr. The volatile componentwas removed in vacuo and the residue was purified with Dowex® 50WX8-100ion-exchange resin (column was washed with water and the compound waseluted with dilute NH₄OH, prepared from 18 ml of NH₄OH and 282 ml ofwater) to afford intermediate (S)-2-amino-4-(diethylamino)butanoic acidas an off-white solid (1.73 g).

Methyl chloroformate (0.36 mL, 4.65 mmol) was added drop-wise over 11min to a cooled (ice-water) mixture of Na₂CO₃ (0.243 g, 2.29 mmol), NaOH(4.6 mL of 1M/H₂O, 4.6 mmol) and the above product (802.4 mg). Thereaction mixture was stirred for 55 min, and then the cooling bath wasremoved and stirring was continued for an additional 5.25 hr. Thereaction mixture was diluted with equal volume of water and washed withCH₂Cl₂ (30 mL, 2×), and the aqueous phase was cooled with ice-water bathand acidified with concentrated HCl to a pH region of 2. The volatilecomponent was then removed in vacuo and the crude material wasfree-based with MCX resin (6.0 g; column was washed with water, andsample was eluted with 2.0 M NH₃/MeOH) to afford impure Cap-76 as anoff-white solid (704 mg). ¹H NMR (MeOH-d₄, δ=3.29 ppm, 400 MHz): δ 3.99(dd, J=7.5, 4.7, 1H), 3.62 (s, 3H), 3.25-3.06 (m, 6H), 2.18-2.09 (m,1H), 2.04-1.96 (m, 1H), 1.28 (t, J=7.3, 6H). LC/MS: Anal. Calcd. for[M+H]⁺ C₁₀H₂₁N₂O₄: 233.15. found 233.24.

Cap-77a and -77b

The synthesis of Cap-77 was conducted according to the proceduredescribed for Cap-7 by using 7-azabicyclo[2.2.1]heptane for the SN₂displacement step, and by effecting the enantiomeric separation of theintermediate benzyl 2-(7-azabicyclo[2.2.1]heptan-7-yl)-2-phenylacetateusing the following condition: the intermediate (303.7 mg) was dissolvedin ethanol, and the resulting solution was injected on a chiral HPLCcolumn (Chiracel AD-H column, 30×250 mm, 5 um) eluting with 90% CO₂-10%EtOH at 70 mL/min, and a temperature of 35° C. to provide 124.5 mg ofenantiomer-1 and 133.8 mg of enantiomer-2. These benzyl esters werehydrogenolysed according to the preparation of Cap-7 to provide Cap-77:¹H NMR (DMSO-d₆, δ=2.5 ppm, 400 MHz): δ 7.55 (m, 2H), 7.38-7.30 (m, 3H),4.16 (s, 1H), 3.54 (app br s, 2H), 2.08-1.88 (m, 4H), 1.57-1.46 (m, 4H).LC (Cond. 1): RT=0.67 min; LC/MS: Anal. Calcd. for [M+H]⁺ C₁₄H₁₈NO₂:232.13. found 232.18. HRMS: Anal. Calcd. for [M+H]⁺ C₁₄H₁₈NO₂: 232.1338.found 232.1340.

Cap-78

NaCNBH₃ (0.5828 g, 9.27 mmol) was added to a mixture of the HCl salt of(R)-2-(ethylamino)-2-phenylacetic acid (an intermediate in the synthesisof Cap-3; 0.9923 mg, 4.60 mmol) and(1-ethoxycyclopropoxy)trimethylsilane (1.640 g, 9.40 mmol) in MeOH (10mL), and the semi-heterogeneous mixture was heated at 50° C. with an oilbath for 20 hr. More (1-ethoxycyclopropoxy)trimethylsilane (150 mg, 0.86mmol) and NaCNBH₃ (52 mg, 0.827 mmol) were added and the reactionmixture was heated for an additional 3.5 hr. It was then allowed to coolto ambient temperature and acidified to a ˜pH region of 2 withconcentrated HCl, and the mixture was filtered and the filtrate wasrotervaped. The resulting crude material was taken up in i-PrOH (6 mL)and heated to effect dissolution, and the non-dissolved part wasfiltered off and the filtrate concentrated in vacuo. About ⅓ of theresultant crude material was purified with a reverse phaseHPLC(H₂O/MeOH/TFA) to afford the TFA salt of Cap-78 as a colorlessviscous oil (353 mg). ¹H NMR (DMSO-d₆, δ=2.5 ppm, 400 MHz; after D₂Oexchange): δ 7.56-7.49 (m, 5H), 5.35 (S, 1H), 3.35 (m, 1H), 3.06 (app brs, 1H), 2.66 (m, 1H), 1.26 (t, J=7.3, 3H), 0.92 (m, 1H), 0.83-0.44 (m,3H). LC (Cond. 1): RT=0.64 min; LC/MS: Anal. Calcd. for [M+H]⁺C₁₃H₁₈NO₂: 220.13. found 220.21. HRMS: Anal. Calcd. for [M+H]⁺C₁₃H₁₈NO₂: 220.1338. found 220.1343.

Cap-79

Ozone was bubbled through a cooled (−78° C.) CH₂Cl₂ (5.0 mL) solutionCap-55 (369 mg, 2.13 mmol) for about 50 min until the reaction mixtureattained a tint of blue color. Me₂S (10 pipet drops) was added, and thereaction mixture was stirred for 35 min. The −78° C. bath was replacedwith a −10° C. bath and stirring continued for an additional 30 min, andthen the volatile component was removed in vacuo to afford a colorlessviscous oil.

NaBH₃CN (149 mg, 2.25 mmol) was added to a MeOH (5.0 mL) solution of theabove crude material and morpholine (500 μL, 5.72 mmol) and the mixturewas stirred at ambient condition for 4 hr. It was cooled to ice-watertemperature and treated with concentrated HCl to bring its pH to ˜2.0,and then stirred for 2.5 hr. The volatile component was removed invacuo, and the residue was purified with a combination of MCX resin(MeOH wash; 2.0 N NH₃/MeOH elution) and a reverse phaseHPLC(H₂O/MeOH/TFA) to afford Cap-79 containing unknown amount ofmorpholine.

In order to consume the morpholine contaminant, the above material wasdissolved in CH₂Cl₂ (1.5 mL) and treated with Et₃N (0.27 mL, 1.94 mmol)followed by acetic anhydride (0.10 mL, 1.06 mmol) and stirred at ambientcondition for 18 hr. THF (1.0 mL) and H₂O (0.5 mL) were added andstirring continued for 1.5 hr. The volatile component was removed invacuo, and the resultant residue was passed through MCX resin (MeOHwash; 2.0 N NH₃/MeOH elution) to afford impure Cap-79 as a brown viscousoil, which was used for the next step without further purification.

Cap-80a and -80b

SOCl₂ (6.60 mL, 90.5 mmol) was added drop-wise over 15 min to a cooled(ice-water) mixture of (S)-3-amino-4-(benzyloxy)-4-oxobutanoic acid(10.04 g, 44.98 mmol) and MeOH (300 mL), the cooling bath was removedand the reaction mixture was stirred at ambient condition for 29 hr.Most of the volatile component was removed in vacuo and the residue wascarefully partitioned between EtOAc (150 mL) and saturated NaHCO₃solution. The aqueous phase was extracted with EtOAc (150 mL, 2×), andthe combined organic phase was dried (MgSO₄), filtered, and concentratedin vacuo to afford (S)-1-benzyl 4-methyl 2-aminosuccinate as a colorlessoil (9.706 g). ¹H NMR (DMSO-d₆, δ=2.5 ppm, 400 MHz): δ 7.40-7.32 (m,5H), 5.11 (s, 2H), 3.72 (app t, J=6.6, 1H), 3.55 (s, 3H), 2.68 (dd,J=15.9, 6.3, 1H), 2.58 (dd, J=15.9, 6.8, 1H), 1.96 (s, 2H). LC (Cond.1): RT=0.90 min; LC/MS: Anal. Calcd. for [M+H]⁺ C₁₂H₁₆NO₄: 238.11. found238.22.

Pb(NO₃)₂ (6.06 g, 18.3 mmol) was added over 1 min to a CH₂Cl₂ (80 mL)solution of (S)-1-benzyl 4-methyl 2-aminosuccinate (4.50 g, 19.0 mmol),9-bromo-9-phenyl-9H-fluorene (6.44 g, 20.0 mmol) and Et₃N (3.0 mL, 21.5mmol), and the heterogeneous mixture was stirred at ambient conditionfor 48 hr. The mixture was filtered and the filtrate was treated withMgSO₄ and filtered again, and the final filtrate was concentrated. Theresulting crude material was submitted to a Biotage purification (350 gsilica gel, CH₂Cl₂ elution) to afford (S)-1-benzyl 4-methyl2-(9-phenyl-9H-fluoren-9-ylamino)succinate as highly viscous colorlessoil (7.93 g). ¹H NMR (DMSO-d₆, δ=2.5 ppm, 400 MHz): δ 7.82 (m, 2H),7.39-7.13 (m, 16H), 4.71 (d, J=12.4, 1H), 4.51 (d, J=12.6, 1H), 3.78 (d,J=9.1, NH), 3.50 (s, 3H), 2.99 (m, 1H), 2.50-2.41 (m, 2H, partiallyoverlapped with solvent). LC (Cond. 1): RT=2.16 min; LC/MS: Anal. Calcd.for [M+H]⁺ C₃₁H₂₈NO₄: 478.20. found 478.19.

LiHMDS (9.2 mL of 1.0 M/THF, 9.2 mmol) was added drop-wise over 10 minto a cooled (−78° C.) THF (50 mL) solution of (S)-1-benzyl 4-methyl2-(9-phenyl-9H-fluoren-9-ylamino)succinate (3.907 g, 8.18 mmol) andstirred for ˜1 hr. MeI (0.57 mL, 9.2 mmol) was added drop-wise over 8min to the mixture, and stirring was continued for 16.5 hr whileallowing the cooling bath to thaw to room temperature. After quenchingwith saturated NH₄Cl solution (5 mL), most of the organic component wasremoved in vacuo and the residue was partitioned between CH₂Cl₂ (100 mL)and water (40 mL). The organic layer was dried (MgSO₄), filtered, andconcentrated in vacuo, and the resulting crude material was purifiedwith a Biotage (350 g silica gel; 25% EtOAc/hexanes) to afford 3.65 g ofa 2S/3S and 2S/3R diastereomeric mixtures of 1-benzyl 4-methyl3-methyl-2-(9-phenyl-9H-fluoren-9-ylamino)succinate in ˜1.0:0.65 ratio(¹H NMR). The stereochemistry of the dominant isomer was not determinedat this juncture, and the mixture was submitted to the next step withoutseparation. Partial ¹H NMR data (DMSO-d₆, δ=2.5 ppm, 400 MHz): majordiastereomer, δ 4.39 (d, J=12.3, 1H of CH₂), 3.33 (s, 3H, overlappedwith H₂O signal), 3.50 (d, J=10.9, NH), 1.13 (d, J=7.1, 3H); minordiastereomer, δ 4.27 (d, J=12.3, 1H of CH₂), 3.76 (d, J=10.9, NH), 3.64(s, 3H), 0.77 (d, J=7.0, 3H). LC (Cond. 1): RT=2.19 min; LC/MS: Anal.Calcd. for [M+H]⁺ C₃₂H₃₀NO₄: 492.22. found 492.15.

Diisobutylaluminum hydride (20.57 ml of 1.0 M in hexanes, 20.57 mmol)was added drop-wise over 10 min to a cooled (−78° C.) THF (120 mL)solution of (2S)-1-benzyl 4-methyl3-methyl-2-(9-phenyl-9H-fluoren-9-ylamino)succinate (3.37 g, 6.86 mmol)prepared above, and stirred at −78° C. for 20 hr. The reaction mixturewas removed from the cooling bath and rapidly poured into ˜1M H₃PO₄/H₂O(250 mL) with stirring, and the mixture was extracted with ether (100mL, 2×). The combined organic phase was washed with brine, dried(MgSO₄), filtered and concentrated in vacuo. A silica gel mesh of thecrude material was prepared and submitted to chromatography (25%EtOAc/hexanes; gravity elution) to afford 1.1 g of (2S,3S)-benzyl4-hydroxy-3-methyl-2-(9-phenyl-9H-fluoren-9-ylamino)butanoate,contaminated with benzyl alcohol, as a colorless viscous oil and(2S,3R)-benzyl4-hydroxy-3-methyl-2-(9-phenyl-9H-fluoren-9-ylamino)butanoate containingthe (2S,3R) stereoisomer as an impurity. The later sample wasresubmitted to the same column chromatography purification conditions toafford 750 mg of purified material as a white foam. [Note: the (2S,3S)isomer elutes before the (2S,3R) isomer under the above condition].(2S,3S) isomer: ¹H NMR (DMSO-d₆, δ=2.5 ppm, 400 MHz): 7.81 (m, 2H),7.39-7.08 (m, 16H), 4.67 (d, J=12.3, 1H), 4.43 (d, J=12.4, 1H), 4.21(app t, J=5.2, OH), 3.22 (d, J=10.1, NH), 3.17 (m, 1H), 3.08 (m, 1H),˜2.5 (m, 1H, overlapped with the solvent signal), 1.58 (m, 1H), 0.88 (d,J=6.8, 3H). LC (Cond. 1): RT=2.00 min; LC/MS: Anal. Calcd. for [M+H]⁺C₃₁H₃₀NO₃: 464.45. found 464.22. (2S,3R) isomer: ¹H NMR (DMSO-d₆, δ=2.5ppm, 400 MHz): 7.81 (d, J=7.5, 2H), 7.39-7.10 (m, 16H), 4.63 (d, J=12.1,1H), 4.50 (app t, J=4.9, 1H), 4.32 (d, J=12.1, 1H), 3.59-3.53 (m, 2H),3.23 (m, 1H), 2.44 (dd, J=9.0, 8.3, 1H), 1.70 (m, 1H), 0.57 (d, J=6.8,3H). LC (Cond. 1): RT=1.92 min; LC/MS: Anal. Calcd. for [M+H]⁺C₃₁H₃₀NO₃: 464.45. found 464.52.

The relative stereochemical assignments of the DIBAL-reduction productswere made based on NOE studies conducted on lactone derivatives preparedfrom each isomer by employing the following protocol: LiHMDS (50 μL of1.0 M/THF, 0.05 mmol) was added to a cooled (ice-water) THF (2.0 mL)solution of (2S,3S)-benzyl4-hydroxy-3-methyl-2-(9-phenyl-9H-fluoren-9-ylamino)butanoate (62.7 mg,0.135 mmol), and the reaction mixture was stirred at similar temperaturefor ˜2 hr. The volatile component was removed in vacuo and the residuewas partitioned between CH₂Cl₂ (30 mL), water (20 mL) and saturatedaqueous NH₄Cl solution (1 mL). The organic layer was dried (MgSO₄),filtered, and concentrated in vacuo, and the resulting crude materialwas submitted to a Biotage purification (40 g silica gel; 10-15%EtOAc/hexanes) to afford(3S,4S)-4-methyl-3-(9-phenyl-9H-fluoren-9-ylamino)dihydrofuran-2(3H)-oneas a colorless film of solid (28.1 mg). (2S,3R)-benzyl4-hydroxy-3-methyl-2-(9-phenyl-9H-fluoren-9-ylamino)butanoate waselaborated similarly to(3S,4R)-4-methyl-3-(9-phenyl-9H-fluoren-9-ylamino)dihydrofuran-2(3H)-one.(3S,4S)-lactone isomer: ¹H NMR (DMSO-d₆, δ=2.5 ppm, 400 MHz), 7.83 (d,J=7.5, 2H), 7.46-7.17 (m, 11H), 4.14 (app t, J=8.3, 1H), 3.60 (d, J=5.8,NH), 3.45 (app t, J=9.2, 1H), ˜2.47 (m, 1H, partially overlapped withsolvent signal), 2.16 (m, 1H), 0.27 (d, J=6.6, 3H). LC (Cond. 1):RT=1.98 min; LC/MS: Anal. Calcd. for [M+Na]⁺ C₂₄H₂₁NNaO₂: 378.15. found378.42. (3S,4R)-lactone isomer: ¹H NMR (DMSO-d₆, δ=2.5 ppm, 400 MHz),7.89 (d, J=7.6, 1H), 7.85 (d, J=7.3, 1H), 7.46-7.20 (m, 11H), 3.95 (dd,J=9.1, 4.8, 1H), 3.76 (d, J=8.8, 1H), 2.96 (d, J=3.0, NH), 2.92 (dd,J=6.8, 3, NCH), 1.55 (m, 1H), 0.97 (d, J=7.0, 3H). LC (Cond. 1): RT=2.03min; LC/MS: Anal. Calcd. for [M+Na]⁺ C₂₄H₂₁NNaO₂: 378.15. found 378.49.

TBDMS-Cl (48 mg, 0.312 mmol) followed by imidazole (28.8 mg, 0.423 mmol)were added to a CH₂Cl₂ (3 ml) solution of (2S,3S)-benzyl4-hydroxy-3-methyl-2-(9-phenyl-9H-fluoren-9-ylamino)butanoate (119.5 mg,0.258 mmol), and the mixture was stirred at ambient condition for 14.25hr. The reaction mixture was then diluted with CH₂Cl₂ (30 mL) and washedwith water (15 mL), and the organic layer was dried (MgSO₄), filtered,and concentrated in vacuo. The resultant crude material was purifiedwith a Biotage (40 g silica gel; 5% EtOAc/hexanes) to afford(2S,3S)-benzyl4-(tert-butyldimethylsilyloxy)-3-methyl-2-(9-phenyl-9H-fluoren-9-ylamino)butanoate,contaminated with TBDMS based impurities, as a colorless viscous oil(124.4 mg). (2S,3R)-benzyl4-hydroxy-3-methyl-2-(9-phenyl-9H-fluoren-9-ylamino)butanoate waselaborated similarly to (2S,3R)-benzyl4-(tert-butyldimethylsilyloxy)-3-methyl-2-(9-phenyl-9H-fluoren-9-ylamino)butanoate.(2S,3S)-silyl ether isomer: ¹H NMR (DMSO-d₆, δ=2.5 ppm, 400 MHz), 7.82(d, J=4.1, 1H), 7.80 (d, J=4.0, 1H), 7.38-7.07 (m, 16H), 4.70 (d,J=12.4, 1H), 4.42 (d, J=12.3, 1H), 3.28-3.19 (m, 3H), 2.56 (dd, J=10.1,5.5, 1H), 1.61 (m, 1H), 0.90 (d, J=6.8, 3H), 0.70 (s, 9H), −0.13 (s,3H), −0.16 (s, 3H). LC (Cond. 1, where the run time was extended to 4min): RT=3.26 min; LC/MS: Anal. Calcd. for [M+H]⁺ C₃₇H₄₄NO₃Si: 578.31.found 578.40. (2S,3R)-silyl ether isomer: ¹H NMR (DMSO-d₆, δ=2.5 ppm,400 MHz), 7.82 (d, J=3.0, 1H), 7.80 (d, J=3.1, 1H), 7.39-7.10 (m, 16H),4.66 (d, J=12.4, 1H), 4.39 (d, J=12.4, 1H), 3.61 (dd, J=9.9, 5.6, 1H),3.45 (d, J=9.5, 1H), 3.41 (dd, J=10, 6.2, 1H), 2.55 (dd, J=9.5, 7.3,1H), 1.74 (m, 1H), 0.77 (s, 9H), 0.61 (d, J=7.1, 3H), −0.06 (s, 3H),−0.08 (s, 3H).

A balloon of hydrogen was attached to a mixture of (2S,3S)-benzyl4-(tert-butyldimethylsilyloxy)-3-methyl-2-(9-phenyl-9H-fluoren-9-ylamino)butanoate(836 mg, 1.447 mmol) and 10% Pd/C (213 mg) in EtOAc (16 mL) and themixture was stirred at room temperature for ˜21 hr, where the balloonwas recharged with H₂ as necessary. The reaction mixture was dilutedwith CH₂Cl₂ and filtered through a pad of diatomaceous earth(Celite-545®), and the pad was washed with EtOAc (200 mL), EtOAc/MeOH(1:1 mixture, 200 mL) and MeOH (750 mL). The combined organic phase wasconcentrated, and a silica gel mesh was prepared from the resultingcrude material and submitted to a flash chromatography (8:2:1 mixture ofEtOAc/i-PrOH/H₂O) to afford(2S,3S)-2-amino-4-(tert-butyldimethylsilyloxy)-3-methylbutanoic acid asa white fluffy solid (325 mg). (2S,3R)-benzyl4-(tert-butyldimethylsilyloxy)-3-methyl-2-(9-phenyl-9H-fluoren-9-ylamino)butanoatewas similarly elaborated to(2S,3R)-2-amino-4-(tert-butyldimethylsilyloxy)-3-methylbutanoic acid.(2S,3S)-amino acid isomer: ¹H NMR (Methanol-d₄, δ=3.29 ppm, 400 MHz),3.76 (dd, J=10.5, 5.2, 1H), 3.73 (d, J=3.0, 1H), 3.67 (dd, J=10.5, 7.0,1H), 2.37 (m, 1H), 0.97 (d, J=7.0, 3H), 0.92 (s, 9H), 0.10 (s, 6H).LC/MS: Anal. Calcd. for [M+H]⁺ C₁₁H₂₆NO₃Si: 248.17. found 248.44.(2S,3R)-amino acid isomer: ¹H NMR (Methanol-d₄, δ=3.29 ppm, 400 MHz),3.76-3.75 (m, 2H), 3.60 (d, J=4.1, 1H), 2.16 (m, 1H), 1.06 (d, J=7.3,3H), 0.91 (s, 9H), 0.09 (s, 6H). Anal. Calcd. for [M+H]⁺ C₁₁H₂₆NO₃Si:248.17. found 248.44.

Water (1 mL) and NaOH (0.18 mL of 1.0 M/H₂O, 0.18 mmol) were added to amixture of(2S,3S)-2-amino-4-(tert-butyldimethylsilyloxy)-3-methylbutanoic acid(41.9 mg, 0.169 mmol) and Na₂CO₃ (11.9 mg, 0.112 mmol), and sonicatedfor about 1 min to effect dissolution of reactants. The mixture was thencooled with an ice-water bath, methyl chloroformate (0.02 mL, 0.259mmol) was added over 30 s, and vigorous stirring was continued atsimilar temperature for 40 min and then at ambient temperature for 2.7hr. The reaction mixture was diluted with water (5 mL), cooled withice-water bath and treated drop-wise with 1.0 N HCl aqueous solution(˜0.23 mL). The mixture was further diluted with water (10 mL) andextracted with CH₂Cl₂ (15 mL, 2×). The combined organic phase was dried(MgSO₄), filtered, and concentrated in vacuo to afford Cap-80a as anoff-white solid.(2S,3R)-2-amino-4-(tert-butyldimethylsilyloxy)-3-methylbutanoic acid wassimilarly elaborated to Cap-80b. Cap-80a: ¹H NMR (DMSO-d₆, δ=2.5 ppm,400 MHz), 12.57 (br s, 1H), 7.64 (d, J=8.3, 0.3H), 7.19 (d, J=8.8,0.7H), 4.44 (dd, J=8.1, 4.6, 0.3H), 4.23 (dd, J=8.7, 4.4, 0.7H),3.56/3.53 (two singlets, 3H), 3.48-3.40 (m, 2H), 2.22-2.10 (m, 1H), 0.85(s, 9H), ˜0.84 (d, 0.9H, overlapped with t-Bu signal), 0.79 (d, J=7,2.1H), 0.02/0.01/0.00 (three overlapping singlets, 6H). LC/MS: Anal.Calcd. for [M+Na]⁺ C₁₃H₂₇NNaO₅Si: 328.16. found 328.46. Cap-80b: ¹H NMR(CDCl₃, δ=7.24 ppm, 400 MHz), 6.00 (br d, J=6.8, 1H), 4.36 (dd, J=7.1,3.1, 1H), 3.87 (dd, J=10.5, 3.0, 1H), 3.67 (s, 3H), 3.58 (dd, J=10.6,4.8, 1H), 2.35 (m, 1H), 1.03 (d, J=7.1, 3H), 0.90 (s, 9H), 0.08 (s, 6H).LC/MS: Anal. Calcd. for [M+Na]⁺ C₁₃H₂₇NNaO₅Si: 328.16. found 328.53. Thecrude products were utilized without further purification.

Cap-81

Prepared according to the protocol described by Falb et al. SyntheticCommunications 1993, 23, 2839.

Cap-82 to Cap-85

Cap-82 to Cap-85 were synthesized from appropriate starting materialsaccording to the procedure described for Cap-51 or Cap-13. The samplesexhibited similar spectral profiles as that of their enantiomers (i.e.,Cap-4, Cap-13, Cap-51 and Cap-52, respectively).

To a mixture of O-methyl-L-threonine (3.0 g, 22.55 mmol), NaOH (0.902 g,22.55 mmol) in H₂O (15 mL) was added ClCO₂Me (1.74 mL, 22.55 mmol)dropwise at 0° C. The mixture was allowed to stir for 12 h and acidifiedto pH 1 using 1N HCl. The aqueous phase was extracted with EtOAc and(2×250 mL) and 10% MeOH in CH₂Cl₂ (250 mL) and the combined organicphases were concentrated under in vacuo to afford a colorless oil (4.18g, 97%) which was of sufficient purity for use in subsequent steps.¹HNMR (400 MHz, CDCl₃) δ 4.19 (s, 1H), 3.92-3.97 (m, 1H), 3.66 (s, 3H),1.17 (d, J=7.7 Hz, 3H). LCMS: Anal. Calcd. for C₇H₁₃NO₅: 191. found: 190(M−H)⁻.

Cap-87

To a mixture of L-homoserine (2.0 g, 9.79 mmol), Na₂CO₃ (2.08 g, 19.59mmol) in H₂O (15 mL) was added ClCO₂Me (0.76 mL, 9.79 mmol) dropwise at0° C. The mixture was allowed to stir for 48 h and acidified to pH 1using 1N HCl. The aqueous phase was extracted with EtOAc and (2×250 mL)and the combined organic phases were concentrated in vacuo to afford acolorless solid (0.719 g, 28%) which was of sufficient purity for use insubsequent steps. ¹HNMR (400 MHz, CDCl₃) δ 4.23 (dd, J=4.5, 9.1 Hz, 1H),3.66 (s, 3H), 3.43-3.49 (m, 2H), 2.08-2.14 (m, 1H), 1.82-1.89 (m, 1H).LCMS: Anal. Calcd. for C₇H₁₃NO₅: 191. found: 192 (M+H)⁺.

Cap-88

A mixture of L-valine (1.0 g, 8.54 mmol), 3-bromopyridine (1.8 mL, 18.7mmol), K₂CO₃ (2.45 g, 17.7 mmol) and CuI (169 mg, 0.887 mmol) in DMSO(10 mL) was heated at 100° C. for 12 h. The reaction mixture was cooledto rt, poured into H₂O (ca. 150 mL) and washed with EtOAc (×2). Theorganic layers were extracted with a small amount of H₂O and thecombined aq phases were acidified to ca. pH 2 with 6N HCl. The volumewas reduced to about one-third and 20 g of cation exchange resin(Strata) was added. The slurry was allowed to stand for 20 min andloaded onto a pad of cation exchange resin (Strata) (ca. 25 g). The padwas washed with H₂O (200 mL), MeOH (200 mL), and then NH₃ (3M in MeOH,2×200 mL). The appropriate fractions was concentrated in vacuo and theresidue (ca. 1.1 g) was dissolved in H₂O, frozen and lyophyllized. Thetitle compound was obtained as a foam (1.02 g, 62%). ¹HNMR (400 MHz,DMSO-d₆) δ 8.00 (s, br, 1H), 7.68-7.71 (m, 1H), 7.01 (s, br, 1H), 6.88(d, J=7.5 Hz, 1H), 5.75 (s, br, 1H), 3.54 (s, 1H), 2.04-2.06 (m, 1H),0.95 (d, J=6.0 Hz, 3H), 0.91 (d, J=6.6 Hz, 3H). LCMS: Anal. Calcd. forC₁₀H₁₄N₂O₂: 194. found: 195 (M+H)⁺.

Cap-89

A mixture of L-valine (1.0 g, 8.54 mmol), 5-bromopyrimidine (4.03 g,17.0 mmol), K₂CO₃ (2.40 g, 17.4 mmol) and CuI (179 mg, 0.94 mmol) inDMSO (10 mL) was heated at 100° C. for 12 h. The reaction mixture wascooled to RT, poured into H₂O (ca. 150 mL) and washed with EtOAc (×2).The organic layers were extracted with a small amount of H₂O and thecombined aq phases were acidified to ca. pH 2 with 6N HCl. The volumewas reduced to about one-third and 20 g of cation exchange resin(Strata) was added. The slurry was allowed to stand for 20 min andloaded onto a pad of cation exchange resin (Strata) (ca. 25 g). The padwas washed with H₂O (200 mL), MeOH (200 mL), and then NH₃ (3M in MeOH,2×200 mL). The appropriate fractions was concentrated in vacuo and theresidue (ca. 1.1 g) was dissolved in H₂O, frozen and lyophyllized. Thetitle compound was obtained as a foam (1.02 g, 62%). ¹HNMR (400 MHz,CD₃OD) showed the mixture to contain valine and the purity could not beestimated. The material was used as is in subsequent reactions. LCMS:Anal. Calcd. for C₉H₁₃N₃O₂: 195. found: 196 (M+H)⁺.

Cap-90

Cap-90 was prepared according to the method described for thepreparation of Cap-1. The crude material was used as is in subsequentsteps. LCMS: Anal. Calcd. for C₁₁H₁₅NO₂: 193. found: 192 (M−H)⁻.

The following caps were prepared according to the method used forpreparation of cap 51 unless noted otherwise:

Cap Structure LCMS Cap-91

LCMS: Anal. Calcd. for C₁₁H₁₃NO₄: 223; found: 222 (M − H)⁻. Cap-92

LCMS: Anal. Calcd. for C₁₁H₁₃NO₄: 223; found: 222 (M − H)⁻. Cap-93

LCMS: Anal. Calcd. for C₁₀H₁₂N₂O₄: 224; found: 225 (M + H)⁺. Cap-94

LCMS: Anal. Calcd. for C₈H₁₁N₃O₄: 224; found: 214 (M + H)⁺. Cap-95

LCMS: Anal. Calcd. for C₁₃H₁₇NO₄: 251; found: 250 (M − H)⁻. Cap-96

LCMS: Anal. Calcd. for C₁₂H₁₅NO₄: 237; found: 236 (M − H)⁻. Cap-97

LCMS: Anal. Calcd. for C₉H₁₅NO₄: 201; found: 200 (M − H)⁻. Cap-98

LCMS: Anal. Calcd. for C₉H₁₅NO₄: 201; found: 202 (M + H)⁺. Cap-99

¹HNMR (400 MHz, CD₃OD) δ 3.88-3.94 (m, 1H), 3.60, 3.61 (s, 3H), 2.80 (m,1H), 2.20 (m 1H), 1.82-1.94 (m, 3H), 1.45- 1.71 (m, 2H). Cap-99a

¹HNMR (400 MHz, CD₃OD) δ 3.88-3.94 (m, 1H), 3.60, 3.61 (s, 3H), 2.80 (m,1H), 2.20 (m 1H), 1.82-1.94 (m, 3H), 1.45- 1.71 (m, 2H). Cap-100

LCMS: Anal. Calcd. for C₁₂H₁₄NO₄F: 255; found: 256 (M + H)⁺. Cap-101

LCMS: Anal. Calcd. for C₁₁H₁₃NO₄: 223; found: 222 (M − H)⁻. Cap-102

LCMS: Anal. Calcd. for C11H13NO4: 223; found: 222 (M − H)⁻ Cap-103

LCMS: Anal. Calcd. for C₁₀H₁₂N₂O₄: 224; found: 225 (M + H)⁺. Cap-104

¹HNMR (400 MHz, CD₃OD) δ 3.60 (s, 3H), 3.50-3.53 (m, 1H), 2.66- 2.69 and2.44-2.49 (m, 1H), 1.91-2.01 (m, 2H), 1.62-1.74 (m, 4H), 1.51- 1.62 (m,2H). Cap-105

¹HNMR (400 MHz, CD₃OD) δ 3.60 (s, 3H), 3.33-3.35 (m, 1H, partiallyobscured by solvent), 2.37-2.41 and 2.16-2.23 (m, 1H), 1.94- 2.01 (m,4H), 1.43- 1.53 (m, 2H), 1.17-1.29 (m, 2H). Cap-106

  Prepared from cis-4- aminocyclohexane carboxylic acid and acetaldehydeby employing a similar procedure described for the synthesis of Cap-2.The crude HCl salt was passed through MCX (MeOH/H₂O/CH₂Cl₂ wash; 2 NNH₃/MeOH elution) to afford an oil. which was dissolved in CH₃CN/H₂O andlyophilized to afford a tan solid. 1HNMR (400 MHz, CD₃OD) δ 3.16 (q, J =7.3 Hz, 4H), 2.38-2.41 (m, 1H), 2.28-2.31 (m, 2H), 1.79-1.89 (m, 2H),1.74 (app. ddd 7 = 3.5, 12.5, 15.9 Hz, 2H), 1.46 (app dt J = 4.0, 12.9Hz, 2H), 1.26 (t, J = 7.3 Hz, 6H) Cap-107

LCMS: Anal. Calcd. for C₈H₁₀N₂O₄S: 230; found: 231 (M + H)⁺. Cap-108

LCMS: Anal. Calcd. for C₁₅H₁₇N₃O₄: 303; found: 304 (M + H)⁺. Cap-109

LCMS: Anal. Calcd. for C₁₀H₁₂N₂O₄: 224; found: 225 (M + H)⁺. Cap-110

LCMS: Anal. Calcd. for C₁₀H₁₂N₂O₄: 224; found: 225 (M + H)⁺. Cap-111

LCMS: Anal. Calcd. for C₁₂H₁₆NO₈P: 333; found: 334 (M + H)⁺. Cap-112

LCMS: Anal. Calcd. for C₁₃H₁₄N₂O₄: 262; found: 263 (M + H)⁺. Cap-113

LCMS: Anal. Calcd. for C₁₈H₁₉NO₅: 329; found: 330 (M + H)⁺. Cap-114

¹HNMR (400 MHz, CDC13) δ 4.82-4.84 (m, 1H), 4.00-4.05 (m, 2H), 3.77 (s,3H), 2.56 (s, br, 2H) Cap-115

¹HNMR (400 MHz, CDCl₃) δ 5.13 (s, br, 1H), 4.13 (s, br, 1H), 3.69 (s, 3H), 2.61 (d, J = 5.0 Hz, 2H), 1.28 (d, J = 9.1 Hz, 3H). Cap-116

¹HNMR (400 MHz, CDCl₃) δ 5.10 (d, J = 8.6 Hz, 1H), 3.74-3.83 (m, 1H),3.69 (s, 3H), 2.54- 2.61 (m, 2H), 1.88 (sept, J = 7.0 Hz, 1H), 0.95 (d,J = 7.0 Hz, 6H).

Cap-117 to Cap-123

For the preparation of Cap-117 to Cap-123 the Boc amino acids wereobtained from commercially sources and were deprotected by treatmentwith 25% TFA in CH₂Cl₂. After complete reaction as judged by LCMS thesolvents were removed in vacuo and the corresponding TFA salt of theamino acid was carbamoylated with methyl chloroformate according to theprocedure described for Cap-51.

Cap Structure LCMS Cap-117

LCMS: Anal. Calcd. for C₁₂H₁₅NO₄: 237; found: 238 (M + H)⁺. Cap-118

LCMS: Anal. Calcd. for C₁₀H₁₃NO₄S: 243; found: 244 (M + H)⁺. Cap-119

LCMS: Anal. Calcd. for C₁₀H₁₃NO₄S: 243; found: 244 (M + H)⁺. Cap-120

LCMS: Anal. Calcd. for C₁₀H₁₃NO₄S: 243; found: 244 (M + H)⁺. Cap-121

¹HNMR (400 MHz, CDCl3) δ 4.06-4.16 (m, 1H), 3.63 (s, 3H), 3.43 (s, 1H),2.82 and 2.66 (s, br, 1H), 1.86- 2.10 (m, 3H), 1.64 - 1.76 (m, 2H),1.44- 1.53 (m, 1H). Cap-122

¹HNMR profile is similar to that of its enantiomer, Cap-121. Cap-123

LCMS: Anal. Calcd. for C₂₇H₂₆N₂O₆: 474; found: 475 (M + H)⁺.

Cap-124

The hydrochloride salt of L-threonine tert-butyl ester was carbamoylatedaccording to the procedure for Cap-51. The crude reaction mixture wasacidified with 1N HCl to pH˜1 and the mixture was extracted with EtOAc(2×50 mL). The combined organic phases were concentrated in vacuo togive a colorless oil which solidified on standing. The aqueous layer wasconcentrated in vacuo and the resulting mixture of product and inorganicsalts was triturated with EtOAc-CH₂Cl₂-MeOH (1:1:0.1) and then theorganic phase concentrated in vacuo to give a colorless oil which wasshown by LCMS to be the desired product. Both crops were combined togive 0.52 g of a solid. ¹HNMR (400 MHz, CD₃OD) δ 4.60 (m, 1H), 4.04 (d,J=5.0 Hz, 1H), 1.49 (d, J=6.3 Hz, 3H). LCMS: Anal. Calcd. for C₅H₇NO₄:145. found: 146 (M+H)⁺.

Cap-125

To a suspension of Pd(OH)₂, (20%, 100 mg), aqueous formaldehyde (37% wt,4 ml), acetic acid, (0.5 mL) in methanol (15 mL) was added(S)-4-amino-2-(tert-butoxycarbonylamino)butanoic acid (1 g, 4.48 mmol).The reaction was purged several times with hydrogen and was stirredovernight with an hydrogen balloon room temp. The reaction mixture wasfiltered through a pad of diatomaceous earth (Celite®), and the volatilecomponent was removed in vacuo. The resulting crude material was used asis for the next step. LC/MS: Anal. Calcd. for C₁₁H₂₂N₂O₄: 246. found:247 (M+H)⁺.

Cap-126

This procedure is a modification of that used to prepare Cap-51. To asuspension of 3-methyl-L-histidine (0.80 g, 4.70 mmol) in THF (10 mL)and H₂O (10 mL) at 0° C. was added NaHCO₃ (0.88 g, 10.5 mmol). Theresulting mixture was treated with ClCO₂Me (0.40 mL, 5.20 mmol) and themixture allowed to stir at 0° C. After stirring for ca. 2 h LCMS showedno starting material remaining. The reaction was acidified to pH 2 with6 N HCl.

The solvents were removed in vacuo and the residue was suspended in 20mL of 20% MeOH in CH₂Cl₂. The mixture was filtered and concentrated togive a light yellow foam (1.21 g,). LCMS and ¹H NMR showed the materialto be a 9:1 mixture of the methyl ester and the desired product. Thismaterial was taken up in THF (10 mL) and H₂O (10 mL), cooled to 0° C.and LiOH (249.1 mg, 10.4 mmol) was added. After stirring ca. 1 h LCMSshowed no ester remaining. Therefore the mixture was acidified with 6NHCl and the solvents removed in vacuo. LCMS and ¹H NMR confirm theabsence of the ester. The title compound was obtained as its HCl saltcontaminated with inorganic salts (1.91 g, >100%). The compound was usedas is in subsequent steps without further purification. ¹HNMR (400 MHz,CD₃OD) δ 8.84, (s, 1H), 7.35 (s, 1H), 4.52 (dd, J=5.0, 9.1 Hz, 1H), 3.89(s, 3H), 3.62 (s, 3H), 3.35 (dd, J=4.5, 15.6 Hz, 1H, partially obscuredby solvent), 3.12 (dd, J=9.0, 15.6 Hz, 1H). LCMS: Anal. Calcd. forC₉H₁₃N₃O₄: 227.09. found: 228.09 (M+H)⁺.

Cap-127

Cap-127 was prepared according to the method for Cap-126 above startingfrom (S)-2-amino-3-(1-methyl-1H-imidazol-4-yl)propanoic acid (1.11 g,6.56 mmol), NaHCO₃ (1.21 g, 14.4 mmol) and ClCO₂Me (0.56 mL, 7.28 mmol).The title compound was obtained as its HCl salt (1.79 g, >100%)contaminated with inorganic salts. LCMS and ¹H NMR showed the presenceof ca. 5% of the methyl ester. The crude mixture was used as is withoutfurther purification. ¹HNMR (400 MHz, CD₃OD) δ 8.90 (s, 1H), 7.35 (s,1H), 4.48 (dd, J=5.0, 8.6 Hz, 1H), 3.89 (s, 3H), 3.62 (s, 3H), 3.35 (m,1H), 3.08 (m, 1H); LCMS: Anal. Calcd. for C₉H₁₃N₃O₄: 227.09. found: 228(M+H)⁺.

Preparation of Cap-128

Step 1. Preparation of (S)-benzyl2-(tert-butoxycarbonylamino)pent-4-ynoate (cj-27b)

cj-27b

To a solution of cj-27a (1.01 g, 4.74 mmol), DMAP (58 mg, 0.475 mmol)and iPr₂NEt (1.7 mL, 9.8 mmol) in CH₂Cl₂ (100 mL) at 0° C. was addedCbz-Cl (0.68 mL, 4.83 mmol). The solution was allowed to stir for 4 h at0° C., washed (1N KHSO₄, brine), dried (Na₂SO₄), filtered, andconcentrated in vacuo. The residue was purified by flash columnchromatography (TLC 6:1 hex:EtOAc) to give the title compound (1.30 g,91%) as a colorless oil. ¹HNMR (400 MHz, CDCl₃) δ 7.35 (s, 5H), 5.35 (d,br, J=8.1 Hz, 1H), 5.23 (d, J=12.2 Hz, 1H), 5.17 (d, J=12.2 Hz, 1H),4.48-4.53 (m, 1H), 2.68-2.81 (m, 2H), 2.00 (t, J=2.5 Hz, 1H), 1.44 (s,9H). LCMS: Anal. Calcd. for C₁₇H₂₁NO₄: 303. found: 304 (M+H)⁺.

Step 2. Preparation of (S)-benzyl3-(1-benzyl-1H-1,2,3-triazol-4-yl)-2-(tert-butoxycarbonylamino)propanoate(cj-28)

cj-28

To a mixture of (S)-benzyl 2-(tert-butoxycarbonylamino)pent-4-ynoate(0.50 g, 1.65 mmol), sodium ascorbate (0.036 g, 0.18 mmol), CuSO₄.5H₂O(0.022 g, 0.09 mmol) and NaN₃ (0.13 g, 2.1 mmol) in DMF-H₂O (5 mL, 4:1)at rt was added BnBr (0.24 mL, 2.02 mmol) and the mixture was warmed to65° C. After 5 h LCMS indicated low conversion. A further portion ofNaN₃ (100 mg) was added and heating was continued for 12 h. The reactionwas poured into EtOAc and H₂O and shaken. The layers were separated andthe aqueous layer extracted 3× with EtOAc and the combined organicphases washed (H₂O x3, brine), dried (Na₂SO₄), filtered, andconcentrated. The residue was purified by flash (Biotage, 40+M 0-5% MeOHin CH₂Cl₂; TLC 3% MeOH in CH₂Cl₂) to afford a light yellow oil whichsolidified on standing (748.3 mg, 104%). The NMR was consistent with thedesired product but suggests the presence of DMF. The material was usedas is without further purification. ¹HNMR (400 MHz, DMSO-d₆) δ 7.84 (s,1H), 7.27-7.32 (m, 10H), 5.54 (s, 2H), 5.07 (s, 2H), 4.25 (m, 1H), 3.16(dd, J=1.0, 5.3 Hz, 1H), 3.06 (dd, J=5.3, 14.7 Hz), 2.96 (dd, J=9.1,14.7 Hz, 1H), 1.31 (s, 9H). LCMS: Anal. Calcd. for C₂₄H₂₈N₄O₄: 436.found: 437 (M+H)⁺.

Step 3. Preparation of (S)-benzyl3-(1-benzyl-1H-1,2,3-triazol-4-yl)-2-(methoxycarbonylamino)propanoate(cj-29)

cj-29

A solution of (S)-benzyl3-(1-benzyl-1H-1,2,3-triazol-4-yl)-2-(tert-butoxycarbonylamino)propanoate(0.52 g, 1.15 mmol) in CH₂Cl₂ was added TFA (4 mL). The mixture wasallowed to stir at room temperature for 2 h. The mixture wasconcentrated in vacuo to give a colorless oil which solidified onstanding. This material was dissolved in THF—H₂O and cooled to 0° C.Solid NaHCO₃ (0.25 g, 3.00 mmol) was added followed by ClCO₂Me (0.25 mL,3.25 mmol). After stirring for 1.5 h the mixture was acidified to pH˜2with 6N HCl and then poured into H₂O-EtOAc. The layers were separatedand the aq phase extracted 2× with EtOAc. The combined org layers werewashed (H₂O, brine), dried (Na₂SO₄), filtered, and concentrated in vacuoto give a colorless oil (505.8 mg, 111%, NMR suggested the presence ofan unidentified impurity) which solidified while standing on the pump.The material was used as is without further purification. ¹HNMR (400MHz, DMSO-d₆) δ 7.87 (s, 1H), 7.70 (d, J=8.1 Hz, 1H), 7.27-7.32 (m,10H), 5.54 (s, 2H), 5.10 (d, J=12.7 Hz, 1H), 5.06 (d, J=12.7 Hz, 1H),4.32-4.37 (m, 1H), 3.49 (s, 3H), 3.09 (dd, J=5.6, 14.7 Hz, 1H), 2.98(dd, J=9.6, 14.7 Hz, 1H). LCMS: Anal. Calcd. for C₂₁H₂₂N₄O₄: 394. found:395 (M+H)⁺.

Step 4. Preparation of(S)-2-(methoxycarbonylamino)-3-(1H-1,2,3-triazol-4-yl)propanoic acid(Cap-128)

Cap-128

(S)-benzyl3-(1-benzyl-1H-1,2,3-triazol-4-yl)-2-(methoxycarbonylamino)propanoate(502 mg, 1.11 mmol) was hydrogenated in the presence of Pd—C (82 mg) inMeOH (5 mL) at atmospheric pressure for 12 h. The mixture was filteredthrough diatomaceous earth (Celite®) and concentrated in vacuo.(S)-2-(methoxycarbonylamino)-3-(1H-1,2,3-triazol-4-yl)propanoic acid wasobtained as a colorless gum (266 mg, 111%) which was contaminated withca. 10% of the methyl ester. The material was used as is without furtherpurification. ¹HNMR (400 MHz, DMSO-d₆) δ 12.78 (s, br, 1H), 7.59 (s,1H), 7.50 (d, J=8.0 Hz, 1H), 4.19-4.24 (m, 1H), 3.49 (s, 3H), 3.12 (dd,J=4.8 Hz, 14.9 Hz, 1H), 2.96 (dd, J=9.9, 15.0 Hz, 1H). LCMS: Anal.Calcd. for C₂H₁₀N₄O₄: 214. found: 215 (M+H)⁺.

Preparation of Cap-129

Step 1. Preparation of(S)-2-(benzyloxycarbonylamino)-3-(1H-pyrazol-1-yl)propanoic acid (cj-31)

cj-31

A suspension of (S)-benzyl 2-oxooxetan-3-ylcarbamate (0.67 g, 3.03mmol), and pyrazole (0.22 g, 3.29 mmol) in CH₃CN (12 mL) was heated at50° C. for 24 h. The mixture was cooled to rt overnight and the solidfiltered to afford(S)-2-(benzyloxycarbonylamino)-3-(1H-pyrazol-1-yl)propanoic acid (330.1mg). The filtrate was concentrated in vacuo and then triturated with asmall amount of CH₃CN (ca. 4 mL) to afford a second crop (43.5 mg).Total yield 370.4 mg (44%). m.p. 165.5-168° C. lit m.p. 168.5-169.5[Vederas et al. J. Am. Chem. Soc. 1985, 107, 7105]. ¹HNMR (400 MHz,CD₃OD) δ 7.51 (d, J=2.0, 1H), 7.48 (s, J=1.5 Hz, 1H), 7.24-7.34 (m, 5H),6.23 m, 1H), 5.05 (d, 12.7H, 1H), 5.03 (d, J=12.7 Hz, 1H), 4.59-4.66 (m,2H), 4.42-4.49 (m, 1H). LCMS: Anal. Calcd. for C₁₄H₁₅N₃O₄: 289. found:290 (M+H)⁺.

Step 2. Preparation of(S)-2-(methoxycarbonylamino)-3-(1H-pyrazol-1-yl)propanoic acid (Cap-129)

cap-129

(S)-2-(benzyloxycarbonylamino)-3-(1H-pyrazol-1-yl)propanoic acid (0.20g, 0.70 mmol) was hydrogenated in the presence of Pd—C (45 mg) in MeOH(5 mL) at atmospheric pressure for 2 h. The product appeared to beinsoluble in MeOH, therefore the reaction mixture was diluted with 5 mLH₂O and a few drops of 6NHCl. The homogeneous solution was filteredthrough diatomaceous earth (Celite®), and the MeOH removed in vacuo. Theremaining solution was frozen and lyophyllized to give a yellow foam(188.9 mg). This material was suspended in THF—H₂O (1:1, 10 mL) and thencooled to 0° C. To the cold mixture was added NaHCO₃ (146.0 mg, 1.74mmol) carefully (evolution of CO₂). After gas evolution had ceased (ca.15 min) ClCO₂Me (0.06 mL, 0.78 mmol) was added dropwise. The mixture wasallowed to stir for 2 h and was acidified to pH˜2 with 6N HCl and pouredinto EtOAc. The layers were separated and the aqueous phase extractedwith EtOAC (×5). The combined organic layers were washed (brine), dried(Na₂SO₄), filtered, and concentrated to give the title compound as acolorless solid (117.8 mg, 79%). ¹HNMR (400 MHz, DMSO-d₆) δ 13.04 (s,1H), 7.63 (d, J=2.6 Hz, 1H), 7.48 (d, J=8.1 Hz, 1H), 7.44 (d, J=1.5 Hz,1H), 6.19 (app t, J=2.0 Hz, 1H), 4.47 (dd, J=3.0, 12.9 Hz, 1H),4.29-4.41 (m, 2H), 3.48 (s, 3H). LCMS: Anal. Calcd. for C₈H₁₁N₃O₄: 213.found: 214 (M+H)⁺.

Cap-130

Cap-130 was prepared by acylation of commercially available(R)-phenylglycine analgous to the procedure given in: Calmes, M.;Daunis, J.; Jacquier, R.; Verducci, J. Tetrahedron, 1987, 43(10), 2285.

Cap-131

Step a

Dimethylcarbamoyl chloride (0.92 mL, 10 mmol) was added slowly to asolution of (S)-benzyl 2-amino-3-methylbutanoate hydrochloride (2.44 g;10 mmol) and Hunig's base (3.67 mL, 21 mmol) in THF (50 mL). Theresulting white suspension was stirred at room temperature overnight (16hours) and concentrated under reduced pressure. The residue waspartitioned between ethyl acetate and water. The organic layer waswashed with brine, dried (MgSO₄), filtered, and concentrated underreduced pressure. The resulting yellow oil was purified by flashchromatography, eluting with ethyl acetate:hexanes (1:1). Collectedfractions were concentrated under vacuum providing 2.35 g (85%) of clearoil. ¹H NMR (300 MHz, DMSO-d₆) δ ppm 0.84 (d, J=6.95 Hz, 3H), 0.89 (d,J=6.59 Hz, 3H), 1.98-2.15 (m, 1H), 2.80 (s, 6H), 5.01-5.09 (m, J=12.44Hz, 1H), 5.13 (d, J=12.44 Hz, 1H), 6.22 (d, J=8.05 Hz, 1H), 7.26-7.42(m, 5H). LC (Cond. 1): RT=1.76 min; MS: Anal. Calcd. for [M+H]⁺C₁₆H₂₂N₂O₃: 279.17. found 279.03.

Step b

To a MeOH (50 mL) solution of the intermediate prepared above (2.35 g;8.45 mmol) was added Pd/C (10%; 200 mg) and the resulting blacksuspension was flushed with N₂ (3×) and placed under 1 atm of H₂. Themixture was stirred at room temperature overnight and filtered though amicrofiber filter to remove the catalyst. The resulting clear solutionwas then concentrated under reduced pressure to obtain 1.43 g (89%) ofCap-131 as a white foam, which was used without further purification. ¹HNMR (500 MHz, DMSO-d₆) δ ppm 0.87 (d, J=4.27 Hz, 3H), 0.88 (d, J=3.97Hz, 3H), 1.93-2.11 (m, 1H), 2.80 (s, 6H), 3.90 (dd, J=8.39, 6.87 Hz,1H), 5.93 (d, J=8.54 Hz, 1H), 12.36 (s, 1H). LC (Cond. 1): RT=0.33 min;MS: Anal. Calcd. for [M+H]⁺ C₈H₁₇N₂O₃: 189.12. found 189.04.

Cap-132

Cap-132 was prepared from (S)-benzyl 2-aminopropanoate hydrochlorideaccording to the method described for Cap-131. ¹H NMR (500 MHz, DMSO-d₆)δ ppm 1.27 (d, J=7.32 Hz, 3H), 2.80 (s, 6H), 4.06 (qt, 1H), 6.36 (d,J=7.32 Hz, 1H), 12.27 (s, 1H). LC (Cond. 1): RT=0.15 min; MS: Anal.Calcd. for [M+H]⁺ C₆H₁₃N₂O₃: 161.09. found 161.00.

Cap-133

Cap-133 was prepared from (S)-tert-butyl 2-amino-3-methylbutanoatehydrochloride and 2-fluoroethyl chloroformate according to the methoddescribed for Cap-47. ¹H NMR (500 MHz, DMSO-d₆) δ ppm 0.87 (t, J=6.71Hz, 6H), 1.97-2.10 (m, 1H), 3.83 (dd, J=8.39, 5.95 Hz, 1H), 4.14-4.18(m, 1H), 4.20-4.25 (m, 1H), 4.50-4.54 (m, 1H), 4.59-4.65 (m, 1H), 7.51(d, J=8.54 Hz, 1H), 12.54 (s, 1H).

Cap-134

Cap-134 was prepared from (S)-diethyl alanine and methyl chloroformateaccording to the method described for Cap-51. ¹H NMR (500 MHz, DMSO-d₆)δ ppm 0.72-0.89 (m, 6H), 1.15-1.38 (m, 4H), 1.54-1.66 (m, 1H), 3.46-3.63(m, 3H), 4.09 (dd, J=8.85, 5.19 Hz, 1H), 7.24 (d, J=8.85 Hz, 1H), 12.55(s, 1H). LC (Cond. 2): RT=0.66 min; LC/MS: Anal. Calcd. for [M+H]⁺C₉H₁₈NO₄: 204.12. found 204.02.

Cap-135

A solution of D-2-amino-(4-fluorophenyl)acetic acid (338 mg, 2.00 mmol),1N HCl in diethylether (2.0 mL, 2.0 mmol) and formalin (37%, 1 mL) inmethanol (5 mL) was subjected to balloon hydrogenation over 10%palladium on carbon (60 mg) for 16 h at 25° C. The mixture was thenfiltered through Celite to afford the HCl salt of Cap-135 as a whitefoam (316 mg, 80%). ¹H NMR (300 MHz, MeOH-d₄) δ 7.59 (dd, J=8.80, 5.10Hz, 2H), 7.29 (t, J=8.6 Hz, 2H), 5.17 (s, 1H), 3.05 (v br s, 3H), 2.63(v br s, 3H); R_(t)=0.19 min (Cond.-MS-W5); 95% homogenity index; LRMS:Anal. Calcd. for [M+H]⁺ C₁₀H₁₃FNO₂: 198.09. found: 198.10.

Cap-136

To a cooled (−50° C.) suspension of 1-benzyl-1H-imidazole (1.58 g, 10.0mmol) in anhydrous diethyl ether (50 mL) under nitrogen was addedn-butyl lithium (2.5 M in hexanes, 4.0 mL, 10.0 mmol) dropwise. Afterbeing stirred for 20 min at −50° C., dry carbon dioxide (passed throughDrierite) was bubbled into the reaction mixture for 10 min before it wasallowed to warm up to 25° C. The heavy precipitate which formed onaddition of carbon dioxide to the reaction mixture was filtered to yielda hygroscopic, white solid which was taken up in water (7 mL), acidifiedto pH=3, cooled, and induced to crystallize with scratching. Filtrationof this precipitate gave a white solid which was suspended in methanol,treated with 1N HCl/diethyl ether (4 mL) and concentrated in vacuo.Lyophilization of the residue from water (5 mL) afforded the HCl salt ofCap-136 as a white solid (817 mg, 40%). ¹H NMR (300 MHz, DMSO-d₆) δ 7.94(d, J=1.5 Hz, 1H), 7.71 (d, J=1.5 Hz, 1H), 7.50-7.31 (m, 5H), 5.77 (s,2H); R_(t)=0.51 min (Cond.-MS-W5); 95% homogenity index; LRMS: Anal.Calc. for [M+H]⁺ C₁₁H₁₂N₂O₂: 203.08. found: 203.11.

Cap-137

Cap-137, step a

A suspension of 1-chloro-3-cyanoisoquinoline (188 mg, 1.00 mmol;prepared according to the procedure in WO 2003/099274) (188 mg, 1.00mmol), cesium fluoride (303.8 mg, 2.00 mmol),bis(tri-tert-butylphosphine)palladium dichloride (10 mg, 0.02 mmol) and2-(tributylstannyl)furan (378 μL, 1.20 mmol) in anhydrous dioxane (10mL) under nitrogen was heated at 80° C. for 16 h before it was cooled to25° C. and treated with saturated, aqueous potassium fluoride solutionwith vigorous stirring for 1 h. The mixture was partitioned betweenethyl acetate and water and the organic phase was separated, washed withbrine, dried over Na₂SO₄, filtered and concentrated. Purification of theresidue on silica gel (elution with 0% to 30% ethyl acetate/hexanes)afforded Cap-137, step a (230 mg, 105%) as a white solid which wascarried forward directly. R_(t)=1.95 min (Cond.-MS-W2); 90% homogeneityindex; LRMS: Anal. Calc. for [M+H]⁺ C₁₄H₈N₂O: 221.07. found: 221.12.

Cap-137

To a suspension of Cap 137, step a, (110 mg, 0.50 mmol) and sodiumperiodate (438 mg, 2.05 mmol) in carbon tetrachloride (1 mL),acetonitrile (1 mL) and water (1.5 mL) was added ruthenium trichloridehydrate (2 mg, 0.011 mmol). The mixture was stirred at 25° C. for 2 hand then partitioned between dichloromethane and water. The aqueouslayer was separated, extracted twice more with dichloromethane and thecombined dichloromethane extracts were dried over Na₂SO₄, filtered andconcentrated. Trituration of the residue with hexanes afforded Cap-137(55 mg, 55%) as a grayish-colored solid. R_(t)=1.10 min (Cond.-MS-W2);90% homogeneity index; LCMS: Anal. Calc. for [M+H]⁺ C₁₁H₈N₂O₂: 200.08.found: 200.08.

Caps 138 to 158

Synthetic Strategy. Method A.

Cap-138

Cap-138, Step a

To a stirred suspension of 5-hydroxyisoquinoline (prepared according tothe procedure in WO 2003/099274) (2.0 g, 13.8 mmol) andtriphenylphosphine (4.3 g, 16.5 mmol) in dry tetrahydrofuran (20 mL) wasadded dry methanol (0.8 mL) and diethyl azodicarboxylate (3.0 mL, 16.5mmol) portionwise. The mixture was stirred at room temperature for 20 hbefore it was diluted with ethyl acetate and washed with brine, driedover Na₂SO₄, filtered and concentrated. The residue was preabsorbed ontosilica gel and chromatographed (elution with 40% ethyl acetate/hexanes)to afford Cap-138, step a (1.00 g, 45%) as a light yellow solid. ¹H NMR(CDCl₃, 500 MHz) δ 9.19 (s, 1H), 8.51 (d, J=6.0 Hz, 1H), 7.99 (d, J=6.0Hz, 1H), 7.52-7.50 (m, 2H), 7.00-6.99 (m, 1H), 4.01 (s, 3H); R_(t)=0.66min (Cond.-D2); 95% homogeneity index; LCMS: Anal. Calc. for [M+H]⁺C₁₀H₁₀NO: 160.08. found 160.1.

Cap-138, Step b

To a stirred solution of Cap 138, step a (2.34 g, 14.7 mmol) inanhydrous dichloromethane (50 mL) at room temperature was addedmeta-chloroperbenzoic acid (77%, 3.42 g, 19.8 mmol) in one portion.After being stirred for 20 h, powdered potassium carbonate (2.0 g) wasadded and the mixture was stirred for 1 h at room temperature before itwas filtered and concentrated in vacuo to afford Cap-138, step b (2.15g, 83%) as a pale, yellow solid which was sufficiently pure to carryforward directly. ¹H NMR (CDCl₃, 400 MHz) δ 8.73 (d, J=1.5 Hz, 1H), 8.11(dd, J=7.3, 1.7 Hz, 1H), 8.04 (d, J=7.1 Hz, 1H), 7.52 (t, J=8.1 Hz, 1H),7.28 (d, J=8.3 Hz, 1H), 6.91 (d, J=7.8 Hz, 1H), 4.00 (s, 3H); R_(t)=0.92min, (Cond.-D1); 90% homogenity index; LCMS: Anal. Calc. for [M+H]⁺C₁₀H₁₀NO₂: 176.07. found: 176.0.

Cap-138, Step c

To a stirred solution of Cap 138, step b (0.70 g, 4.00 mmol) andtriethylamine (1.1 mL, 8.00 mmol) in dry acetonitrile (20 mL) at roomtemperature under nitrogen was added trimethylsilylcyanide (1.60 mL,12.00 mmol). The mixture was heated at 75° C. for 20 h before it wascooled to room temperature, diluted with ethyl acetate and washed withsaturated sodium bicarbonate solution and brine prior to drying overNa₂SO₄ and solvent concentration. The residue was flash chromatographedon silica gel (gradient elution with 5% ethyl acetate in hexanes to 25%ethyl acetate in hexanes) to afford Cap-138, step c (498.7 mg, 68%) as awhite, crystalline solid along with 223 mg (30%) of additional Cap-138,step c recovered from the filtrate. ¹H NMR (CDCl₃, 500 MHz) δ 8.63 (d,J=5.5 Hz, 1H), 8.26 (d, J=5.5 Hz, 1H), 7.88 (d, J=8.5 Hz, 1H), 7.69 (t,J=8.0 Hz, 1H), 7.08 (d, J=7.5 Hz, 1H), 4.04 (s, 3H); R_(t)=1.75 min,(Cond.-D1); 90% homogeneity index; LCMS: Anal. Calc. for [M+H]⁺C₁₁H₉N₂O: 185.07. found: 185.10.

Cap-138

Cap-138, step c (0.45 g, 2.44 mmol) was treated with 5N sodium hydroxidesolution (10 mL) and the resulting suspension was heated at 85° C. for 4h, cooled to 25° C., diluted with dichloromethane and acidified with 1Nhydrochloric acid. The organic phase was separated, washed with brine,dried over Na₂SO₄, concentrated to ¼ volume and filtered to affordCap-138 (0.44 g, 88.9%) as a yellow solid. ¹H NMR (DMSO-d₆, 400 MHz) δ13.6 (br s, 1H), 8.56 (d, J=6.0 Hz, 1H), 8.16 (d, J=6.0 Hz, 1H), 8.06(d, J=8.8 Hz, 1H), 7.71-7.67 (m, 1H), 7.30 (d, J=8.0 Hz, 1H), 4.02 (s,3H); R_(t)=0.70 min (Cond.-D1); 95% homogenity index; LCMS: Anal. Calc.for [M+H]⁺ C₁₁H₁₀NO₃: 204.07. found: 204.05.

Synthetic Strategy. Method B (derived from Tetrahedron Letters, 2001,42, 6707).

Cap-139

Cap-139, Step a

To a thick-walled, screw-top vial containing an argon-degassedsuspension of 1-chloro-6-methoxyisoquinoline (1.2 g, 6.2 mmol; preparedaccording to the procedure in WO 2003/099274), potassium cyanide (0.40g, 6.2 mmol), 1,5-bis(diphenylphosphino)pentane (0.27 g, 0.62 mmol) andpalladium (II) acetate (70 mg, 0.31 mmol) in anhydrous toluene (6 mL)was added N,N,N′,N′-tetramethylethylenediamine (0.29 mL, 2.48 mmol). Thevial was sealed, heated at 150° C. for 22 h and then allowed to cool to25° C. The reaction mixture was diluted with ethyl acetate, washed withwater and brine, dried over Na₂SO₄, filtered and concentrated. Theresidue was purified on silica gel (gradient elution with 5% ethylacetate/hexanes to 25% ethyl acetate/hexanes) to afford Cap-139, step a(669.7 mg, 59%) as a white solid. ¹H NMR (CDCl₃, 500 MHz) δ 8.54 (d,J=6.0 Hz, 1H), 8.22 (d, J=9.0 Hz, 1H), 7.76 (d, J=5.5 Hz, 1H), 7.41-7.39(m, 1H), 7.13 (d, J=2.0 Hz, 1H), 3.98 (s, 3H); R_(t)=1.66 min(Cond.-D1); 90% homogenity index; LCMS: Anal. Calc. for [M+H]⁺ C₁₁H₉N₂O:185.07. found: 185.2.

Cap-139

Cap-139 was prepared from the basic hydrolysis of Cap-139, step a with5N

NaOH according to the procedure described for Cap 138. ¹H NMR (400 MHz,DMSO-d₆) δ 13.63 (v br s, 1H), 8.60 (d, J=9.3 Hz, 1H), 8.45 (d, J=5.6Hz, 1H), 7.95 (d, J=5.9 Hz, 1H), 7.49 (d, J=2.2 Hz, 1H), 7.44 (dd,J=9.3, 2.5 Hz, 1H), 3.95 (s, 3H); R_(t)=0.64 min (Cond.-D1); 90%homogenity index; LCMS: Anal. Calc. for [M+H]⁺ C₁₁H₁₀NO₃: 204.07. found:204.05.

Cap-140

Cap-140, Step a

To a vigorously-stirred mixture of 1,3-dichloro-5-ethoxyisoquinoline(482 mg, 2.00 mmol; prepared according to the procedure in WO2005/051410), palladium (II) acetate (9 mg, 0.04 mmol), sodium carbonate(223 mg, 2.10 mmol) and 1,5-bis(diphenylphosphino)pentane (35 mg, 0.08mmol) in dry dimethylacetamide (2 mL) at 25° C. under nitrogen was addedN,N,N′,N′-tetramethylethylenediamine (60 mL, 0.40 mmol). After 10 min,the mixture was heated to 150° C., and then a stock solution of acetonecyanohydrin (prepared from 457 μL of acetone cyanohydrin in 4.34 mL DMA)was added in 1 mL portions over 18 h using a syringe pump. The mixturewas then partitioned between ethyl acetate and water and the organiclayer was separated, washed with brine, dried over Na₂SO₄, filtered andconcentrated. The residue was purified on silica gel (gradient elutionwith 10% ethyl acetate in hexanes to 40% ethyl acetate in hexanes) toafford Cap-140, step a (160 mg, 34%) as a yellow solid. R_(t)=2.46 min(Cond.-MS-W2); 90% homogenity index; LCMS: Anal. Calc. for [M+H]⁺C₁₂H₉ClN₂O: 233.05. found: 233.08.

Cap-140

Cap-140 was prepared by the acid hydrolysis of Cap-140, step a with 12NHCl as described in the procedure for the preparation of Cap 141,described below. R_(t)=2.24 min (Cond.-MS-W2); 90% homogenity index;LCMS: Anal. Calc. for [M+H]⁺ C₁₂H₁₁ClNO₃: 252.04. found: 252.02.

Cap-141

Cap-141, Step a

Cap-141, step a was prepared from 1-bromo-3-fluoroisoquinoline (preparedfrom 3-amino-1-bromoisoquinoline using the procedure outlined in J. Med.Chem. 1970, 13, 613) as described in the procedure for the preparationof Cap-140, step a (vide supra). ¹H NMR (500 MHz, CDCl₃) δ 8.35 (d,J=8.5 Hz, 1H), 7.93 (d, J=8.5 Hz, 1H), 7.83 (t, J=7.63 Hz, 1H),7.77-7.73 (m, 1H), 7.55 (s, 1H); R_(t)=1.60 min (Cond.-D1); 90%homogenity index; LCMS: Anal. Calc. for [M+H]⁺ C₁₀H₆FN₂: 173.05. found:172.99.

Cap-141

Cap-141, step a (83 mg, 0.48 mmol) was treated with 12N HCl (3 mL) andthe resulting slurry was heated at 80° C. for 16 h before it was cooledto room temperature and diluted with water (3 mL). The mixture wasstirred for 10 min and then filtered to afford Cap-141 (44.1 mg, 48%) asan off-white solid. The filtrate was diluted with dichloromethane andwashed with brine, dried over Na₂SO₄, and concentrated to affordadditional Cap-141 (29.30 mg, 32%) which was sufficiently pure to becarried forward directly. ¹H NMR (DMSO-d₆, 500 MHz) δ 14.0 (br s, 1H),8.59-8.57 (m, 1H), 8.10 (d, J=8.5 Hz, 1H), 7.88-7.85 (m, 2H), 7.74-7.71(m, 1H); R_(t)=1.33 min (Cond.-D1); 90% homogenity index; LCMS: Anal.Calc. for [M+H]⁺ C₁₀H₇FNO₂: 192.05. found: 191.97.

Cap-142

Cap-142, Step a

Cap-142, step a was prepared from 4-bromoisoquinoline N-oxide asdescribed in the two-step procedure for the preparation of Cap-138,steps b and c. R_(t)=1.45 min (Cond.-MS-W1); 90% homogenity index; LCMS:Anal. Calc. for [M+H]⁺ C₁₀H₆BrN₂: 232.97. found: 233.00.

Cap-142, Step b

To an argon-degassed suspension of Cap-142, step a (116 mg, 0.50 mmol),potassium phosphate tribasic (170 mg, 0.80 mmol), palladium (II) acetate(3.4 mg, 0.015 mmol) and 2-(dicyclohexylphosphino)biphenyl (11 mg, 0.03mmol) in anhydrous toluene (1 mL) was added morpholine (61 μL, 0.70mmol). The mixture was heated at 100° C. for 16 h, cooled to 25° C.,filtered through diatomaceous earth (Celite®) and concentrated.Purification of the residue on silica gel (gradient elution with 10% to70% ethyl acetate in hexanes) afforded Cap-142, step b (38 mg, 32%) as ayellow solid which was carried forward directly. R_(t)=1.26 min(Cond.-MS-W1); 90% homogenity index; LCMS: Anal. Calc. for [M+H]⁺C₁₄H₁₄N₃O: 240.11. found: 240.13.

Cap-142

Cap-142 was prepared from Cap-142, step b with 5N sodium hydroxide asdescribed in the procedure for Cap 138. R_(t)=0.72 min (Cond.-MS-W1);90% homogenity index; LCMS: Anal. Calc. for [M+H]⁺ C₁₄H₁₅N₂O₃: 259.11.found: 259.08.

Cap-143

Cap-143, Step a

To a stirred solution of 3-amino-1-bromoisoquinoline (444 mg, 2.00 mmol)in anhydrous dimethylformamide (10 mL) was added sodium hydride (60%,unwashed, 96 mg, 2.4 mmol) in one portion. The mixture was stirred at25° C. for 5 min before 2-bromoethyl ether (90%, 250 μL, 2.00 mmol) wasadded. This mixture was stirred further at 25° C. for 5 h and at 75° C.for 72 h before it was cooled to 25° C., quenched with saturatedammonium chloride solution and diluted with ethyl acetate. The organiclayer was separated, washed with water and brine, dried over Na₂SO₄,filtered and concentrated. Purification of the residue on silica gel(gradient elution with 0% to 70% ethyl acetate in hexanes) affordedCap-143, step a (180 mg, 31%) as a yellow solid. R_(t)=1.75 min(Cond.-MS-W1); 90% homogenity index; LCMS: Anal. Calc. for [M+H]⁺C₁₃H₁₄BrN₂O: 293.03. found: 293.04.

Cap-143

To a cold (−60° C.) solution of Cap-143, step a (154 mg, 0.527 mmol) inanhydrous tetrahydrofuran (5 mL) was added a solution of n-butyllithiumin hexanes (2.5 M, 0.25 mL, 0.633 mmol). After 10 min, dry carbondioxide was bubbled into the reaction mixture for 10 min before it wasquenched with 1N HCl and allowed to warm to 25° C. The mixture was thenextracted with dichloromethane (3×30 mL) and the combined organicextracts were concentrated in vacuo. Purification of the residue byreverse phase HPLC (MeOH/water/TFA) afforded Cap-143 (16 mg, 12%).R_(t)=1.10 min (Cond.-MS-W1); 90% homogenity index; LCMS: Anal. Calc.for [M+H]⁺ C₁₄H₁₅N₂O₃: 259.11. found: 259.08.

Cap-144

Cap-144, Step a

1,3-Dichloroisoquinoline (2.75 g, 13.89 mmol) was added in smallportions to a cold (0° C.) solution of fuming nitric acid (10 mL) andconcentrated sulfuric acid (10 mL). The mixture was stirred at 0° C. for0.5 h before it was gradually warmed to 25° C. where it stirred for 16h. The mixture was then poured into a beaker containing chopped ice andwater and the resulting suspension was stirred for 1 h at 0° C. beforeit was filtered to afford Cap-144, step a (2.73 g, 81%) as a yellowsolid which was used directly. R_(t)=2.01 min (Cond.-D1); 95% homogenityindex; LCMS: Anal. Calc. for [M+H]⁺ C₉H₅Cl₂N₂O₂: 242.97. found: 242.92.

Cap-144, Step b

Cap-144, step a (0.30 g, 1.23 mmol) was taken up in methanol (60 mL) andtreated with platinum oxide (30 mg), and the suspension was subjected toParr hydrogenation at 7 psi H₂ for 1.5 h before formalin (5 mL) andadditional platinum oxide (30 mg) were added. The suspension wasresubjected to Parr hydrogenation at 45 psi H₂ for 13 h before it wassuction-filtered through diatomaceous earth (Celite®) and concentrateddown to ¼ volume. Suction-filtration of the ensuing precipitate affordedthe title compound as a yellow solid which was flash chromatographed onsilica gel (gradient elution with 5% ethyl acetate in hexanes to 25%ethyl acetate in hexanes) to afford Cap-144, step b (231 mg, 78%) as apale, yellow solid. R_(t)=2.36 min (Cond.-D1); 95% homogenity index; ¹HNMR (400 MHz, CDCl₃) δ 8.02 (s, 1H), 7.95 (d, J=8.6 Hz, 1H), 7.57-7.53(m, 1H), 7.30 (d, J=7.3 Hz, 1H), 2.88 (s, 6H); LCMS: Anal. Calc. for[M+H]⁺ C₁₁H₁₁Cl₂N₂: 241.03. found: 241.02. HRMS: Anal. Calc. for [M+H]⁺C₁₁H₁₁Cl₂N₂: 241.0299. found: 241.0296.

Cap-144, Step c

Cap-144, step c was prepared from Cap-144, step b according to theprocedure described for the preparation of Cap-139, step a. R_(t)=2.19min (Cond.-D1); 95% homogenity index; LCMS: Anal. Calc. for [M+H]⁺C₁₂H₁₁ClN₃: 232.06. found: 232.03. HRMS: Anal. Calc. for [M+H]⁺C₁₂H₁₁ClN₃: 232.0642. found: 232.0631.

Cap-144

Cap-144 was prepared according to the procedure described for Cap-141.R_(t)=2.36 min (Cond.-D1); 90%; LCMS: Anal. Calc. for [M+H]⁺C₁₂H₁₂ClN₂O₂: 238.01. found: 238.09.

Caps-145 to -162

Caps-145 to 162 were prepared from the appropriate 1-chloroisoquinolinesaccording to the procedure described for the preparation of Cap-138(Method A) or Cap-139 (Method B) unless noted otherwise as outlinedbelow.

Rt (LC-Cond.); % homogeneity index; Cap # Cap Method Hydrolysis MS dataCap-145

  Prepared from commercially available 1,3- dichloroisoquinoline B 12NHCl 1.14 min (Cond.- MS-W1); 90%; LCMS: Anal. Calc. for [M + H]⁺C₁₀H₇ClNO₂: 208.02; found: 208.00. Cap-146

  Prepared from commercially available 3- hydroxyisoquinoline A 5N NaOH1.40 min (Cond.- D1); 95%; LCMS: Anal. Calc. for [M + H]⁺ C₁₁H₁₀NO₃:204.07; found: 204.06. Cap-147

  Prepared from commercially available 1- chloro-4-hydroxyisoquinoline B5N NaOH 0.87 min (Cond.- D1); 95%; LCMS: Anal. Calc. for [M + H]⁺C₁₁H₁₀NO₃: 204.07; found: 204.05. Cap-148

  Prepared from commercially available 7- hydroxyisoquinoline A 5N NaOH0.70 min (Cond- D1); 95%; LCMS: Anal. Calc. for [M + H]⁺ C₁₁H₁₀NO₃:204.07; found: 204.05. Cap-149

  Prepared from commercially available 5- hydroxyisoquinoline A 5N NaOH0.70 min (Cond.- D1); 95%; LCMS: Anal. Calc. for [M + H]⁺ C₁₁H₁₀NO₃:204.07; found: 204.05. Cap-150

  Prepared from 8-methoxy- 1-chloroisoquinoline, which can besynthesized following the procedure in WO 2003/099274 A 12N HCl 0.26 min(Cond.- D1); 95%; LCMS: Anal. Calc. for [M + H]⁺ C₁₁H₁₀NO₃: 204.07;found: 204.04. Cap-151

  Prepared from 5-methoxy- 1,3-dichloroisoquinoline, which can besynthesized following the procedure in WO 2005/051410. B 12N HCl 1.78min (Cond.- D1); 90%; LCMS: Anal. Calc. for [M + H]⁺ C₁₁H₉ClNO₃: 238.03;found: 238.09. Cap-152

  Prepared from commercially available 6- methoxy-1,3-dichloroisoquinoline B 12N HCl 1.65 min (Cond.- D1); 95%; LCMS: Anal.Calc. for [M + H]⁺ C₁₁H₉ClNO₃: 238.00; found: 238.09. Cap-153

  Prepared from 4- bromoisoquinoline, which can be synthesized followingthe procedure in WO 2003/062241 A 6N HCl 1.18 min (Cond.- MS-W1); 95%;LCMS: Anal. Calc. for [M + H]⁺ C₁₀H₇BrNO₂: 251.97; found: 251.95.Cap-154

  Prepared from 7-fluoro-1- chloroisoquinoline, which can be synthesizedfollowing the procedure in WO 2003/099274 B 5N NaOH 0.28 min (Cond.-MS-W1); 90%; LCMS: Anal. Calc. for [M + H]⁺ C₁₀H₇FNO₂: 192.05; found:192.03. Cap-155

  Prepared from 1,7- dichloroisoquinoline, which can be synthesizedfollowing the procedure in WO 2003/099274 B 5N NaOH 0.59 min (Cond.-MS-W1); 90%; LCMS: Anal. Calc. for [M + H]⁺ C₁₀H₇ClNO₂: 208.02; found:208.00. Cap-156

  Prepared from 1,6- dichloroisoquinoline, which can be synthesizedfollowing the procedure in WO 2003/099274 B 5N NaOH 0.60 min (Cond.-MS-W1); 90%; LCMS: Anal. Calc. for [M + H]⁺ C₁₀H₇ClNO₂: 208.02; found:208.03. Cap-157

  Prepared from 1,4- dichloroisoquinoline, which can be synthesizedfollowing the procedure in WO 2003/062241 B 1.49 min (Cond.- Dl); 95%;LCMS: Anal. Calc. for [M + H]⁺ C₁₀H₁₇ClNO: 208.02; found: 208.00.Cap-158

  Prepared from 1,5- dichloroisoquinoline, which can be synthesizedfollowing the procedure in WO 2003/099274 B 5N NaOH 0.69 min (Cond.-MS-W1); 90%; LCMS: Anal. Calc. for [M + H]⁺ C₁₀H₇ClNO₂: 208.02; found:208.01. Cap-159

  Prepared from 5-fluoro-1- chloroisoquinoline, which can be synthesizedfollowing the procedure in WO 2003/099274 B 5N NaOH 0.41 min (Cond.-MS-W1); 90%; LCMS: Anal. Calc. for [M + H]⁺ C₁₀H₇FNO₂: 192.05; found:192.03. Cap-160

  Prepared from 6-fluoro-1- chloroisoquinoline, which can be synthesizedfollowing the procedure in WO 2003/099274 B 5N NaOH 0.30 min (Cond.-MS-W1); 90%; LCMS: Anal. Calc. for [M + H]⁺ C₁₀H₇FNO₂: 192.05; found:192.03. Cap-161

  Prepared from 4- bromoquinoline-2-carboxylic acid and dimethylamine(DMSO, 100° C.) — — 0.70 min (Cond. D1); 95%; LCMS: Anal. Calc. for [M +H]⁺ C₁₂H₁₃N₂O₂: 217.10; found: 217.06. Cap-162

  Prepared from m-anisidine following the procedure described in J.Hetero. Chem. 1993, 17 and Heterocycles, 2003, 60, 953. — — 0.65 min(Cond.- M3); 95%; LCMS: Anal. Calc. for [M + H]⁺ C₁₁H₁₀NO₃: 204.07;found: 203.94.

Cap-163

To a solution of 2-ketobutyric acid (1.0 g, 9.8 mmol) in diethylether(25 ml) was added phenylmagnesium bromide (22 ml, 1M in THF) dropwise.The reaction was stirred at ˜25° C. under nitrogen for 17.5 h. Thereaction was acidified with 1N HCl and the product was extracted withethyl acetate (3×100 ml). The combined organic layer was washed withwater followed by brine and dried over MgSO₄. After concentration invacuo, a white solid was obtained. The solid was recrystallized fromhexanes/ethyl acetate to afford Cap-163 as white needles (883.5 mg). ¹HNMR (DMSO-d₆, δ=2.5 ppm, 500 MHz): 12.71 (br s, 1H), 7.54-7.52 (m, 2H),7.34-7.31 (m, 2H), 7.26-7.23 (m, 1H), 5.52-5.39 (br s, 1H), 2.11 (m,1H), 1.88 (m, 1H), 0.79 (app t, J=7.4 Hz, 3H).

Cap-164

A mixture of 2-amino-2-phenylbutyric acid (1.5 g, 8.4 mmol),formaldehyde (14 mL, 37% in water), 1N HCl (10 mL) and 10% Pd/C (0.5 mg)in MeOH (40 mL) was exposed to H₂ at 50 psi in a Parr bottle for 42 h.The reaction was filtered over Celite and concentrated in vacuo, theresidue was taken up in MeOH (36 mL) and the product was purified with areverse phase HPLC (MeOH/H₂O/TFA) to afford the TFA salt of Cap-164 as awhite solid (1.7 g). ¹H NMR (DMSO-d₆, δ=2.5 ppm, 500 MHz) 7.54-7.47 (m,5H), 2.63 (m, 1H), 2.55 (s, 6H), 2.31 (m, 1H), 0.95 (app t, J=7.3 Hz,3H).

Cap-165

To a mixture of 2-amino-2-indanecarboxylic acid (258.6 mg, 1.46 mmol)and formic acid (0.6 ml, 15.9 mmol) in 1,2-dichloroethane (7 ml) wasadded formaldehyde (0.6 ml, 37% in water). The mixture was stirred at˜25° C. for 15 min then heated at 70° C. for 8 h. The volatile componentwas removed in vacuo, and the residue was dissolved in DMF (14 mL) andpurified by a reverse phase HPLC (MeOH/H₂O/TFA) to afford the TFA saltof Cap-165 as a viscous oil (120.2 mg). ¹H NMR (DMSO-d₆, δ=2.5 ppm, 500MHz): 7.29-7.21 (m, 4H), 3.61 (d, J=17.4 Hz, 2H), 3.50 (d, J=17.4 Hz,2H), 2.75 (s, 6H). LC/MS: Anal. Calcd. for [M+H]⁺ C₁₂H₁₆NO₂: 206.12.found: 206.07.

Cap-166a and -166b

Caps-166a and -166b were prepared from(1S,4S)-(+)-2-methyl-2,5-diazabicyclo[2.2.1]heptane (2HBr) according tothe method described for the synthesis of Cap-7a and Cap-7b, with theexception that the benzyl ester intermediate was separated using asemi-prep Chrialcel OJ column, 20×250 mm, 10 μm eluting with 85:15heptane/ethanol mixture at 10 mL/min elution rate for 25 min. Cap-166b:¹H NMR (DMSO-d₆, δ=2.5 ppm, 500 MHz): 7.45 (d, J=7.3 Hz, 2H), 7.27-7.19(m, 3H), 4.09 (s, 1H), 3.34 (app br s, 1H), 3.16 (app br s, 1H), 2.83(d, J=10.1 Hz, 1H), 2.71 (m, 2H), 2.46 (m, 1H), 2.27 (s, 3H), 1.77 (d,J=9.8 Hz, 1H), 1.63 (d, J=9.8 Hz, 1H). LC/MS: Anal. Calcd. for [M+H]⁺C₁₄H₁₉N₂O₂: 247.14. found: 247.11.

Cap-167

A solution of racemic Boc-1,3-dihydro-2H-isoindole carboxylic acid (1.0g, 3.8 mmol) in 20% TFA/CH₂Cl₂ was stirred at ˜25° C. for 4 h. All thevolatile component was removed in vacuo. A mixture of the resultantcrude material, formaldehyde (15 mL, 37% in water), 1N HCl (10 mL) and10% Pd/C (10 mg) in MeOH was exposed to H₂ (40 PSI) in a Parr bottle for23 h. The reaction mixture was filtered over Celite and concentrated invacuo to afford Cap-167 as a yellow foam (873.5 mg). ¹H NMR (DMSO-d₆,δ=2.5 ppm, 500 MHz) 7.59-7.38 (m, 4H), 5.59 (s, 1H), 4.84 (d, J=14 Hz,1H), 4.50 (d, J=14.1 Hz, 1H), 3.07 (s, 3H). LC/MS: Anal. Calcd. for[M+H]⁺ C₁₀H₁₂NO₂: 178.09. found: 178.65.

Cap-168

Racemic Cap-168 was prepared from racemic Boc-aminoindane-1-carboxylicacid according to the procedure described for the preparation ofCap-167. The crude material was employed as such.

Cap-169

A mixture of 2-amino-2-phenylpropanoic acid hydrochloride (5.0 g, 2.5mmol), formaldehyde (15 ml, 37% in water), 1N HCl (15 ml), and 10% Pd/C(1.32 g) in MeOH (60 mL) was placed in a Parr bottle and shaken underhydrogen (55 PSI) for 4 days. The reaction mixture was filtered overCelite and concentrated in vacuo. The residue was taken up in MeOH andpurified by reverse phase prep-HPLC (MeOH/water/TFA) to afford the TFAsalt of Cap-169 as a viscous semi-solid (2.1 g). ¹H NMR (CDCl₃, δ=7.26ppm, 500 MHz): 7.58-7.52 (m, 2H), 7.39-7.33 (m, 3H), 2.86 (br s, 3H),2.47 (br s, 3H), 1.93 (s, 3H). LC/MS: Anal. Calcd. for [M+H]⁺ C₁₁H₁₆NO₂:194.12. found: 194.12.

Cap-170

To (S)-2-amino-2-(tetrahydro-2H-pyran-4-yl)acetic acid (505 mg; 3.18mmol; obtained from Astatech) in water (15 ml) was added sodiumcarbonate (673 mg; 6.35 mmol), and the resultant mixture was cooled to0° C. and then methyl chloroformate (0.26 ml; 3.33 mmol) was addeddropwise over 5 minutes. The reaction was allowed to stir for 18 hourswhile allowing the bath to thaw to ambient temperature. The reactionmixture was then partitioned between 1N HCl and ethyl acetate. Theorganic layer was removed and the aqueous layer was further extractedwith 2 additional portions of ethyl acetate. The combined organic layerswere washed with brine, dried over magnesium sulfate, filtered andconcentrated in vacuo to afford Cap-170a colorless residue. ¹H NMR (500MHz, DMSO-d₆) δ ppm 12.65 (1H, br s), 7.44 (1H, d, J=8.24 Hz), 3.77-3.95(3H, m), 3.54 (3H, s), 3.11-3.26 (2H, m), 1.82-1.95 (1H, m), 1.41-1.55(2H, m), 1.21-1.39 (2H, m); LC/MS: Anal. Calcd. for [M+H]⁺ C₉H₁₆NO₅:218.1. found 218.1.

Cap-171

A solution of methyl2-(benzyloxycarbonylamino)-2-(oxetan-3-ylidene)acetate (200 mg, 0.721mmol; I1 Farmaco (2001), 56, 609-613) in ethyl acetate (7 ml) and CH₂Cl₂(4.00 ml) was degassed by bubbling nitrogen for 10 min. Dimethyldicarbonate (0.116 ml, 1.082 mmol) and Pd/C (20 mg, 0.019 mmol) werethen added, the reaction mixture was fitted with a hydrogen balloon andallowed to stir at ambient temperature overnight at which time TLC (95:5CH₂Cl₂/MeOH: visulalized with stain made from 1 g Ce(NH₄)₂SO₄, 6 gammonium molybdate, 6 ml sulfuric acid, and 100 ml water) indicatedcomplete conversion. The reaction was filtered through celite andconcentrated. The residue was purified via Biotage® (load withdichloromethane on 25 samplet; elute on 25S column with dichloromethanefor 3CV then 0 to 5% MeOH/dichloromethane over 250 ml then hold at 5%MeOH/dichloromethane for 250 ml; 9 ml fractions). Collected fractionscontaining desired material and concentrated to 120 mg (81%) of methyl2-(methoxycarbonylamino)-2-(oxetan-3-yl)acetate as a colorless oil. ¹HNMR (500 MHz, CHLOROFORM-D) δ ppm 3.29-3.40 (m, J=6.71 Hz, 1H) 3.70 (s,3H) 3.74 (s, 3H) 4.55 (t, J=6.41 Hz, 1H) 4.58-4.68 (m, 2H) 4.67-4.78 (m,2H) 5.31 (br s, 1H). LC/MS: Anal. Calcd. for [M+H]⁺ C₈H₁₄NO₅: 204.2.found 204.0.

To methyl 2-(methoxycarbonylamino)-2-(oxetan-3-yl)acetate (50 mg, 0.246mmol) in THF (2 mL) and water (0.5 mL) was added lithium hydroxidemonohydrate (10.33 mg, 0.246 mmol). The resultant solution was allowedto stir overnite at ambient temperature. TLC (1:1 EA/Hex; Hanessianstain [1 g Ce(NH₄)₂SO₄, 6 g ammonium molybdate, 6 ml sulfuric acid, and100 ml water]) indicated ˜10% starting material remaining. Added anadditional 3 mg LiOH and allowed to stir overnight at which time TLCshowed no starting material remaining. Concentrated in vacuo and placedon high vac overnite providing 55 mg lithium2-(methoxycarbonylamino)-2-(oxetan-3-yl)acetate as a colorless solid. ¹HNMR (500 MHz, MeOD) δ ppm 3.39-3.47 (m, 1H) 3.67 (s, 3H) 4.28 (d, J=7.93Hz, 1H) 4.64 (t, J=6.26 Hz, 1H) 4.68 (t, J=7.02 Hz, 1H) 4.73 (d, J=7.63Hz, 2H).

Cap-172

Cap-172, Step a

The following diazotization step was adapted from Barton, A.;Breukelman, S. P.; Kaye, P. T.; Meakins, G. D.; Morgan, D. J. J. C. S.Perkin Trans I 1982, 159-164: A solution of NaNO₂ (166 mg, 2.4 mmol) inwater (0.6 mL) was added slowly to a stirred, cold (0° C.) solution ofmethyl 2-amino-5-ethyl-1,3-thiazole-4-carboxylate (186 mg, 1.0 mmol),CuSO₄.5H₂O (330 mg, 1.32 mmol), NaCl (260 mg, 4.45 mmol) and H₂SO₄ (5.5mL) in water (7.5 mL). The mixture was stirred at 0° C. for 45 min andallowed to warm up to room temperature where it stirred further for 1 hbefore CuCl (118 mg) was added. This mixture was stirred further at roomtemperature for 16 h before it was diluted with brine and extracted withether twice. The organic layers were combined, dried over MgSO₄ andconcentrated to give methyl 2-chloro-5-ethylthiazole-4-carboxylate (i.e.Cap-172, step a) (175 mg, 85%) as an orange oil (80% pure) which wasused directly in the next reaction. R_(t)=1.99 min (Cond.-MD1); LC/MS:Anal. Calcd. for [M+H]⁺ C₇H₉ClNO₂S: 206.01. found: 206.05.

Cap-172

To a solution of methyl 2-chloro-5-ethylthiazole-4-carboxylate (175 mg)in THF/H₂O/MeOH (20 mL/3 mL/12 mL) was added LiOH (305 mg, 12.76 mmol).The mixture was stirred at room temperature overnight before it wasconcentrated down and neutralized with 1N HCl in ether (25 mL). Theresidue was extracted twice with ethyl acetate and the organic layerswere combined, dried over MgSO₄ and evaporated to yield Cap-172 (60 mg,74%) as a red solid which was used without further purification. ¹H NMR(300 MHz, DMSO-d₆) δ ppm 13.03-13.42 (1H, m), 3.16 (2H, q, J=7.4 Hz),1.23 (3H, t, J=7.5 Hz). R_(t)=1.78 min (Cond.-MD1); LC/MS: Anal. Calcd.for [M+H]⁺ C₆H₇ClNO₂S: 191.99. found: 191.99.

Cap-173

Cap-173, Step a

The following diazotization step was adapted from Barton, A.;Breukelman, S. P.; Kaye, P. T.; Meakins, G. D.; Morgan, D. J. J. C. S.Perkin Trans I 1982, 159-164: A solution of NaNO₂ (150 mg, 2.17 mmol) inwater (1.0 mL) was added dropwise to a stirred, cold (0° C.) solution ofmethyl 2-amino-5-ethyl-1,3-thiazole-4-carboxylate (186 mg, 1.0 mmol) in50% H₃PO₂ (3.2 mL). The mixture was stirred at 0° C. for 1 h and allowedto warm up to room temperature where it stirred further for 2 h. Afterrecooling to 0° C., the mixture was treated slowly with a solution ofNaOH (85 mg) in water (10 mL). The mixture was then diluted withsaturated NaHCO₃ solution and extracted twice with ether. The organiclayers were combined, dried over MgSO₄ and concentrated to give methyl5-ethylthiazole-4-carboxylate (i.e. Cap-173, step a) (134 mg, 78%) as anorange oil (85% pure) which was used directly in the next reaction.R_(t)=1.58 min (Cond.-MD1); LC/MS: Anal. Calcd. for [M+H]⁺ C₂H₁₀NO₂S:172.05. found: 172.05.

Cap-173

To a solution of methyl 5-ethylthiazole-4-carboxylate (134 mg) inTHF/H₂O/MeOH (18 mL/2.7 mL/11 mL) was added LiOH (281 mg, 11.74 mmol).The mixture was stirred at room temperature overnight before it wasconcentrated down and neutralized with 1N HCl in ether (25 mL). Theresidue was extracted twice with ethyl acetate and the organic layerswere combined, dried over MgSO₄ and evaporated to yield Cap-173 (90 mg,73%) as an orange solid which was used without further purification. ¹HNMR (300 MHz, DMSO-d₆) δ ppm 12.74-13.04 (1H, m), 3.20 (2H, q, J=7.3Hz), 1.25 (3H, t, J=7.5 Hz). R_(t)=1.27 min (Cond.-MD1); LC/MS: Anal.Calcd. for [M+H]⁺ C₆H₈NO₂S: 158.03. found: 158.04.

Cap-174

Cap-174, Step a

Triflic anhydride (5.0 g, 18.0 mmol) was added dropwise to a cold (0°C.) solution of methyl 3-hydroxypicolinate (2.5 g, 16.3 mmol) and TEA(2.5 mL, 18.0 mmol) in CH₂Cl₂ (80 mL). The mixture was stirred at 0° C.for 1 h before it was allowed to warm up to room temperature where itstirred for an additional 1 h. The mixture was then quenched withsaturated NaHCO₃ solution (40 mL) and the organic layer was separated,washed with brine, dried over MgSO₄ and concentrated to give methyl3-(trifluoromethylsulfonyloxy)picolinate (i.e. Cap-174, step a) (3.38 g,73%) as a dark brown oil (>95% pure) which was used directly withoutfurther purification. ¹H NMR (300 MHz, CDCl₃) δ ppm 8.72-8.79 (1H, m),7.71 (1H, d, J=1.5 Hz), 7.58-7.65 (1H, m), 4.04 (3H, s). R_(t)=1.93 min(Cond.-MD1); LC/MS: Anal. Calcd. for [M+H]⁺ C₈H₇F₃NO₅S: 286.00. found:286.08.

Cap-174

To a solution of methyl 3-(trifluoromethylsulfonyloxy)picolinate (570mg, 2.0 mmol) in DMF (20 mL) was added LiCl (254 mg, 6.0 mmol),tributyl(vinyl)stannane (761 mg, 2.4 mmol) andbis(triphenylphosphine)palladium dichloride (42 mg, 0.06 mmol). Themixture was heated at 100° C. overnight before a saturated solution ofKF (20 mL) was added to the reaction mixture at room temperature. Thismixture was stirred for 4 h before it was filtered through Celite andthe pad of Celite was washed with ethyl acetate. The aqueous phase ofthe filtrate was then separated and concentrated down in vacuo. Theresidue was treated with 4N HCl in dioxanes (5 mL) and the resultingmixture was extracted with methanol, filtered and evaporated to affordCap-174 (260 mg) as a green solid which was slightly contaminated withinorganic salts but was used without further purification. ¹H NMR (300MHz, DMSO-d₆) δ ppm 8.21 (1H, d, J=3.7 Hz), 7.81-7.90 (1H, m), 7.09 (1H,dd, J=7.7, 4.8 Hz), 6.98 (1H, dd, J=17.9, 11.3 Hz), 5.74 (1H, dd,J=17.9, 1.5 Hz), 5.20 (1H, d, J=11.0 Hz). R_(t)=0.39 min (Cond.-MD1);LC/MS: Anal. Calcd. for [M+H]⁺ C₈H₈NO₂: 150.06. found: 150.07.

Cap-175

Cap-175, Step a

To a solution of methyl 3-(trifluoromethylsulfonyloxy)picolinate (i.e.Cap 173, step a) (570 mg, 2.0 mmol), an intermediate in the preparationof Cap-174, in DMF (20 mL) was added LiCl (254 mg, 6.0 mmol),tributyl(vinyl)stannane (761 mg, 2.4 mmol) andbis(triphenylphosphine)palladium dichloride (42 mg, 0.06 mmol). Themixture was heated at 100° C. for 4 h before the solvent was removed invacuo. The residue was taken up in acetonitrile (50 mL) and hexanes (50mL) and the resulting mixture was washed twice with hexanes. Theacetonitrile layer was then separated, filtered through Celite, andevaporated. Purification of the residue by flash chromatography on aHorizon instrument (gradient elution with 25% ethyl acetate in hexanesto 65% ethyl acetate in hexanes) afforded methyl 3-vinylpicolinate (i.e.Cap-175, step a) (130 mg, 40%) as a yellow oil. ¹H NMR (300 MHz, CDCl₃)δ ppm 8.60 (1H, dd, J=4.6, 1.7 Hz), 7.94 (1H, d, J=7.7 Hz), 7.33-7.51(2H, m), 5.72 (1H, d, J=17.2 Hz), 5.47 (1H, d, J=11.0 Hz), 3.99 (3H, s).R_(t)=1.29 min (Cond.-MD1); LC/MS: Anal. Calcd. for [M+H]⁺ C₉H₁₀NO₂:164.07. found: 164.06.

Cap-175, Step b

Palladium on carbon (10%, 25 mg) was added to a solution of methyl3-vinylpicolinate (120 mg, 0.74 mmol) in ethanol (10 mL). The suspensionwas stirred at room temperature under an atmosphere of hydrogen for 1 hbefore it was filtered through Celite and the pad of Celite was washedwith methanol. The filtrate was concentrated down to dryness to yieldmethyl 3-ethylpicolinate (i.e. Cap-175, step b) which was taken directlyinto the next reaction. R_(t)=1.15 min (Cond.-MD1); LC/MS: Anal. Calcd.for [M+H]⁺ C₉H₁₂NO₂: 166.09. found: 166.09.

Cap-175

To a solution of methyl 3-ethylpicolinate in THF/H₂O/MeOH (5 mL/0.75mL/3 mL) was added LiOH (35 mg, 1.47 mmol). The mixture was stirred atroom temperature for 2 d before additional LiOH (80 mg) was added. Afteran additional 24 h at room temperature, the mixture was filtered and thesolvent was removed in vacuo. The residue was then treated with 4N HClin dioxanes (5 mL) and the resulting suspension was concentrated down todryness to yield Cap-175 as a yellow solid which was used withoutfurther purification. ¹H NMR (300 MHz, DMSO-d₆) δ ppm 8.47 (1H, dd,J=4.8, 1.5 Hz), 7.82-7.89 (1H, m), 7.53 (1H, dd, J=7.7, 4.8 Hz), 2.82(2H, q, J=7.3 Hz), 1.17 (3H, t, J=7.5 Hz). R_(t)=0.36 min (Cond.-MD1);LC/MS: Anal. Calcd. for [M+H]⁺ C₈H₁₀NO₂: 152.07. found: 152.10.

Cap-176

Cap-176, Step a

A solution of 1,4-dioxaspiro[4.5]decan-8-one (15 g, 96 mmol) in EtOAc(150 mL) was added to a solution of methyl2-(benzyloxycarbonylamino)-2-(dimethoxyphosphoryl)acetate (21.21 g, 64.0mmol) in 1,1,3,3-tetramethylguanidine (10.45 mL, 83 mmol) and EtOAc (150mL). The resulting solution was the stirred at ambient temperature for72 h and then it was diluted with EtOAc (25 mL). The organic layer waswashed with 1N HCl (75 mL), H2O (100 mL) and brine (100 mL), dried(MgSO₄), filtered and concentrated. The residue was purified via Biotage(5% to 25% EtOAc/Hexanes; 300 g column). The combined fractionscontaining the product were then concentrated under vacuum and theresidue was re-crystallized from hexanes/EtOAc to give white crystalsthat corresponded to methyl2-(benzyloxycarbonylamino)-2-(1,4-dioxaspiro[4.5]decan-8-ylidene)acetate(6.2 g) ¹H NMR (400 MHz, CDCl₃-d) δ ppm 7.30-7.44 (5H, m), 6.02 (1H, br.s.), 5.15 (2H, s), 3.97 (4H, s), 3.76 (3H, br. s.), 2.84-2.92 (2H, m),2.47 (2H, t, J=6.40 Hz), 1.74-1.83 (4H, m). LC (Cond. OL1): R_(t)=2.89min. LC/MS: Anal. Calcd. For [M+Na]⁺ C₁₉H₂₃NNaO₆: 745.21. found: 745.47.

Cap 176, Step b

Ester Cap 176, step b was prepared from alkene Cap 176, step a accordingto the method of Burk, M. J.; Gross, M. F. and Martinez J. P. (J. Am.Chem. Soc., 1995, 117, 9375-9376 and references therein): A 500 mLhigh-pressure bottle was charged with alkene Cap 176, step a (3.5 g,9.68 mmol) in degassed MeOH (200 mL) under a blanket of N₂. The solutionwas then charged with(−)-1,2-Bis((2S,5S)-2,5-dimethylphospholano)ethane(cyclooctadiene)rhodium(I) tetrafluoroborate (0.108 g, 0.194 mmol) and the resulting mixturewas flushed with N₂ (3×) and charged with H₂ (3×). The solution wasshaken vigorously under 70 psi of H₂ at ambient temperature for 72 h.The solvent was removed under reduced pressure and the remaining residuewas taken up in EtOAc. The brownish solution was then filtered through aplug of Silica Gel and eluted with EtOAc. The solvent was concentratedunder vacuum to afford a clear oil corresponding to ester Cap 176, stepb (3.4 g). ¹H NMR (500 MHz, CDCl₃-d) δ ppm 7.28-7.43 (5H, m), 5.32 (1H,d, J=9.16 Hz), 5.06-5.16 (2H, m), 4.37 (1H, dd, J=9.00, 5.04 Hz), 3.92(4H, t, J=3.05 Hz), 3.75 (3H, s), 1.64-1.92 (4 H, m), 1.37-1.60 (5H, m).LC (Cond. OL1): R_(t)=1.95 min. LC/MS: Anal. Calcd. For [M+H]⁺C₁₉H₂₆NO₆: 364.18. found: 364.27.

Cap 176, Step c

Ester Cap 176, step b (4.78 g, 13.15 mmol) was dissolved in THF (15 mL)followed by sequential addition of water (10 mL), glacial acetic acid(26.4 mL, 460 mmol) and dichloroacetic acid (5.44 mL, 65.8 mmol). Theresulting mixture was stirred for 72 h at ambient temperature, and thereaction was quenched by slow addition of solid Na₂CO₃ with vigorousstirring until the release of gas was no longer visible. Crude productwas extracted into 10% ethyl acetate-dichloromethane and the organiclayers were combined, dried (MgSO₄) filtered and concentrated. Theresulting residue was purified via Biotage (0 to 30% EtOAc/Hex; 25 gcolumn) to afford ketone Cap 176, step c (3.86 g) as a clear oil. ¹H NMR(400 MHz, CDCl₃-d) δ ppm 7.28-7.41 (5H, m), 5.55 (1H, d, J=8.28 Hz),5.09 (2H, s), 4.46 (1H, dd, J=8.16, 5.14 Hz), 3.74 (3H, s), 2.18-2.46(5H, m), 1.96-2.06 (1H, m), 1.90 (1H, ddd, J=12.99, 5.96, 2.89 Hz),1.44-1.68 (2H, m, J=12.36, 12.36, 12.36, 12.36, 4.77 Hz). LC (Cond.OL1): R_(t)=1.66 min. LC/MS: Anal. Calcd. For [M+Na]⁺ C₁₇H₂₁NNaO₅:342.13. found: 342.10.

Cap 176, Step d

Deoxo-Fluor® (3.13 mL, 16.97 mmol) was added to a solution of ketone Cap176, step c (2.71 g, 8.49 mmol) in CH₂Cl₂ (50 mL) followed by additionof a catalytic amount of EtOH (0.149 mL, 2.55 mmol). The resultingyellowish solution was stirred at rt overnight. The reaction wasquenched by addition of sat. aq. NaHCO₃ (25 mL) and the mixture wasextracted with EtOAc (3×75 mL)). The combined organic layers were dried(MgSO₄), filtered and dried to give a yellowish oil. The residue waspurified via Biotage chromatography (2% to 15% EtOAc/Hex; 90 g column)and a white solid corresponding to the difluoro amino acid dilforide Cap176, step d (1.5 g) was recovered. ¹H NMR (400 MHz, CDCl₃-d) δ ppm7.29-7.46 (5H, m), 5.34 (1H, d, J=8.28 Hz), 5.12 (2H, s), 4.41 (1H, dd,J=8.66, 4.89 Hz), 3.77 (3H, s), 2.06-2.20 (2H, m), 1.83-1.98 (1H, m),1.60-1.81 (4H, m), 1.38-1.55 (2H, m). ¹⁹F NMR (376 MHz, CDCl₃-d) δ ppm−92.15 (1F, d, J=237.55 Hz), −102.44 (1F, d, J=235.82 Hz). LC (Cond.OL1): R_(t)=1.66 min. LC/MS: Anal. Calcd. For [2M+Na]⁺ C₃₄H₄₂F₄N₂NaO₈:705.28. found: 705.18.

Cap 176, Step e

Difluoride Cap 176, step d (4 g, 11.72 mmol) was dissolved in MeOH (120mL) and charged with Pd/C (1.247 g, 1.172 mmol). The suspension wasflushed with N₂ (3×) and the reaction mixture was placed under 1 atm ofH₂ (balloon). The mixture was stirred at ambient temperature for 48 h.The suspension was then filtered though a plug of Celite andconcentrated under vacuum to give an oil that corresponded to amino acidCap 176, step e (2.04 g) and that was used without further purification.¹H NMR (400 MHz, DMSO-d₆) δ ppm 3.62 (3H, s), 3.20 (1H, d, J=5.77 Hz),1.91-2.09 (2H, m), 1.50-1.88 (7H, m), 1.20-1.45 (2H, m). ¹⁹F NMR (376MHz, DMSO-d₆) δ ppm −89.39 (1F, d, J=232.35 Hz), −100.07 (1F, d,J=232.35 Hz). ¹³C NMR (101 MHz, DMSO-d₆) δ ppm 175.51 (1C, s), 124.10(1C, t, J=241.21, 238.90 Hz), 57.74 (1C, s), 51.39 (1C, s), 39.23 (1C,br. s.), 32.02-33.83 (2C, m), 25.36 (1C, d, J=10.02 Hz), 23.74 (1C, d,J=9.25 Hz). LC (Cond. OL2): R_(t)=0.95 min. LC/MS: Anal. Calcd. For[2M+H]⁺ C₁₈H₃₁F₄N₂O₂: 415.22. found: 415.40.

Cap 176, Step f

Methyl chloroformate (1.495 mL, 19.30 mmol) was added to a solution ofamino acid Cap 176, step e (2 g, 9.65 mmol) and DIEA (6.74 mL, 38.6mmol) in CH₂Cl₂ (100 mL). The resulting solution was stirred at rt for 3h and volatiles were removed under reduced pressure. The residue waspurified via Biotage (0% to 20% EtOAc/Hex; 90 g column). A clear oilthat solidified upon standing under vacuum and corresponding tocarbamate Cap-176, step f (2.22 g) was recovered. ¹H NMR (500 MHz,CDCl₃-d) δ ppm 5.27 (1H, d, J=8.55 Hz), 4.39 (1H, dd, J=8.85, 4.88 Hz),3.77 (3H, s), 3.70 (3H, s), 2.07-2.20 (2H, m), 1.84-1.96 (1H, m),1.64-1.82 (4H, m), 1.39-1.51 (2H, m). ¹⁹F NMR (471 MHz, CDCl₃-d) δ ppm−92.55 (1F, d, J=237.13 Hz), −102.93 (1F, d, J=237.12 Hz). ¹³C NMR (126MHz, CDCl₃-d) δ ppm 171.97 (1C, s), 156.69 (1C, s), 119.77-125.59 (1C,m), 57.24 (1C, br. s.), 52.48 (1C, br. s.), 52.43 (1C, s), 39.15 (1C,s), 32.50-33.48 (2C, m), 25.30 (1C, d, J=9.60 Hz), 24.03 (1C, d, J=9.60Hz). LC (Cond. OL1): R_(t)=1.49 min. LC/MS: Anal. Calcd. For [M+Na]⁺C₁₁H₁₇F₂NNaO₄: 288.10. found: 288.03.

Cap-176

A solution of LiOH (0.379 g, 15.83 mmol) in Water (25 mL) was added to asolution of carbamate Cap-176, step f (2.1 g, 7.92 mmol) in THF (75 mL)and the resulting mixture was stirred at ambient temperature for 4 h.THF was removed under vacuum and the remaining aqueous phase wasacidified with 1N HCl solution (2 mL) and then extracted with EtOAc(2×50 mL). The combined organic layers were dried (MgSO₄), filtered andconcentrated to give a white foam corresponding to Cap-176 (1.92 g). ¹HNMR (400 MHz, DMSO-d₆) δ ppm 12.73 (1H, s), 7.50 (1H, d, J=8.78 Hz),3.97 (1H, dd, J=8.53, 6.02 Hz), 3.54 (3H, s), 1.92-2.08 (2H, m),1.57-1.90 (5H, m), 1.34-1.48 (1H, m), 1.27 (1H, qd, J=12.72, 3.26 Hz).¹⁹F NMR (376 MHz, DMSO-d₆) δ ppm −89.62 (1F, d, J=232.35 Hz), −99.93(1F, d, J=232.35 Hz). LC (Cond. OL2): R_(t)=0.76 min. LC/MS: Anal.Calcd. For [M−H]⁺ C₁₀H₁₄F₂NO₄: 250.09. found: 250.10.

EXAMPLES

The present disclosure will now be described in connection with certainembodiments which are not intended to limit its scope. On the contrary,the present disclosure covers all alternatives, modifications, andequivalents as can be included within the scope of the claims. Thus, thefollowing examples, which include specific embodiments, will illustrateone practice of the present disclosure, it being understood that theexamples are for the purposes of illustration of certain embodiments andare presented to provide what is believed to be the most useful andreadily understood description of its procedures and conceptual aspects.

Solution percentages express a weight to volume relationship, andsolution ratios express a volume to volume relationship, unless statedotherwise. Nuclear magnetic resonance (NMR) spectra were recorded eitheron a Bruker 300, 400, or 500 MHz spectrometer; the chemical shifts (6)are reported in parts per million.

Purity assessment and low resolution mass analysis were conducted on aShimadzu LC system coupled with Waters Micromass ZQ MS system. It shouldbe noted that retention times may vary slightly between machines. Unlessnoted otherwise, the LC conditions employed in determining the retentiontime (R_(t)) were:

Condition 1

Start % B=0

Final % B=100

Gradient time=3 min

Stop time=4 min

Flow Rate=4 mL/min

Wavelength=220 nm

Solvent A=0.1% TFA in 10% methanol/90% water

Solvent B=0.1% TFA in 90% methanol/10% water

Column=PHENOMENEX® 10u C18 3.0×50 mm

Condition 2

Start % B=0

Final % B=100

Gradient time=2 min

Stop time=3 min

Flow rate=4 mL/min

Wavelength=220 nm

Solvent A=10% MeOH/90% H₂O/0.1% TFA

Solvent B=90% MeOH/10% H₂O/0.1% TFA

Column 2=PHENOMENEX®-Luna 3.0×50 mm S10

Condition 2a

Start % B=0

Final % B=100

Gradient time=3 min

Stop time=4 min

Flow Rate=4 mL/min

Wavelength=220 nm

Solvent A=0.1% TFA in 10% methanol/90% water

Solvent B=0.1% TFA in 90% methanol/10% water

Column=PHENOMENEX®-Luna 3.0×50 mm S10

Condition 3

Start % B=0

Final % B=100

Gradient time=3 min

Stop time=4 min

Flow Rate=4 mL/min

Wavelength=220 nm

Solvent A=0.1% TFA in 10% methanol/90% water

Solvent B=0.1% TFA in 90% methanol/10% water

Column=PHENOMENEX®-Luna C18 10u 3.0×50 mm

Condition 4

Start % B=0

Final % B=100

Gradient time=3 min

Stop time=4 min

Flow Rate=0.8 mL/min

Wavelength=220 nm

Solvent A=0.1% TFA in 10% methanol/90% water

Solvent B=0.1% TFA in 90% methanol/10% water

Column=PHENOMENEX®-Luna 2.0×50 mm, 3 μm

Oven Temp.=40° C.

Example QC-1

Example QC-1a

To a solution of bicyclohexanone (5.05 g, 26.0 mmol) and 2,6-diterbutylphenol (11.75 g, 57.2 mmol) in 200 mL of dry dichloromethane was addedtriflic anhydride (9.10 mL, 54.6 mmol). The resulting solution wasstirred at room temperature overnight. The solvent was removed undervacuum. The residue was taken up in hexanes and filtered. The filtratewas washed with 1 N HCl and brine. The organic layer was dried withpotassium carbonate and concentrated. The crude product was purified bya flash chromatography (silica gel, 5% ethyl acetate/hexanes) to giveExample QC-1a as a white solid (8.22 g, 69.0%, mixture ofdiastereomers). ¹H NMR (500 MHz, CDCl₃) δ ppm 1.35-1.64 (m, 4H)1.81-2.07 (m, 4H) 2.10-2.53 (m, 6H) 5.66-5.80 (m, 2H). LC/MS: Anal.Calcd. for [M+H]⁺ C₁₄H₁₇F₆O₆S₂: 459.37. found (molecule did not ionizewell in mass spec chamber).

Example QC-1b

To a flask with bis(pinacolato)diboron (2.79 g, 11.0 mmol), potassiumphenolate (1.98 g, 15.0 mmol), PdCl₂(PPh₃)₂ (0.21 g, 0.3 mmol) andtriphenylphosphine (0.16 g, 0.6 mmol) was added 50 mL of dry toluene and(1a) (2.29 g, 5.0 mmol). The resulting mixture was stirred at 50° C. for3 h. The reaction was quenched with water and extracted with toluene.The organic layers were combined, washed with brine, dried with MgSO₄and concentrated. The crude product was purified by flash chromatography(silica gel, 5-80% ethyl acetate/hexanes) to give Example QC-1b as awhite solid (1.14 g, 55%, mixture of diastereomers). ¹H NMR (500 MHz,CDCl₃) δ ppm 1.10-1.19 (m, 2H) 1.24 (s, 24H) 1.31-1.43 (m, 2H) 1.73-1.90(m, 4H) 1.97-2.29 (m, 6H) 6.52-6.58 (m, 2H).

Example QC-1c

Glyoxal (11 mL of 40% in water) was added drop-wise over 11 min to amethanol (32 mL) solution of ammonium hydroxide (32 mL) and(S)-Boc-prolinal (8.564 g, 42.98 mmol) and stirred at ambienttemperature for 19 h. The volatile component was removed in vacuo andthe residue was purified by a flash chromatography (silica gel, ethylacetate) followed by a recrystallization (ethyl acetate, roomtemperature) to afford imidazole Example QC-1c as a white fluffy solid(4.43 g). ¹H NMR (DMSO-D₆, δ=2.50, 400 MHz): 11.68/11.59 (br s, 1H),6.94 (s, 1H), 6.76 (s, 1H), 4.76 (m, 1H), 3.48 (m, 1H), 3.35-3.29 (m,1H), 2.23-1.73 (m, 4H), 1.39/1.15 (two s, 9H). LC (Cond. 2): RT=0.87min; >95% homogeneity index. LC/MS: Anal. Calcd. for [M+H]⁺ C₁₂H₂₀N₃O₂238.16. found 238.22.

Imidazole Example QC-1c had an ee of 98.9% when analyzed under chiralHPLC condition noted below:

Column: Chiralpak AD, 10 um, 4.6×50 mm

Solvent: 1.7% ethanol/heptane (isocratic)

Flow rate: 1 ml/min

Wavelength: either 220 or 256 nm

Relative retention time: 3.25 min (R), 5.78 min (5)

Example QC-1d

To a solution of Example QC-1c (4.98 g, 21.0 mmol) in 35 mL of drydimethylformamide was added sodium hydride (0.88 g of 60% in mineraloil, 22.0 mmol). The resulting mixture was stirred at room temperaturefor 40 min. 2-(trimethylsilyl)ethoxymethyl chloride (3.75 mL, 21.0 mmol)was then added dropwise over 5 min. The resulting solution was stirredat room temperature for 2 h. The reaction was quenched with water, andextracted with ethyl acetate. The organic layers were combined, washedwith brine, dried with MgSO₄ and concentrated. The crude product waspurified by flash chromatography (silica gel, 35-50% ethylacetate/hexanes) to give Example QC-1d as a colorless oil (6.30 g,81.6%). ¹H NMR (500 MHz, CDCl₃) δ ppm −0.03 (s, 9H) 0.76-0.99 (m, 2H)1.20/1.38 (rotomers, s/s, 9H) 1.76-2.50 (m, 4H) 3.37-3.77 (m, 4H)4.80-5.03 (m, 1H) 5.07-5.16 (m, Hz, 1H) 5.40/5.78 (rotomers, d/d,J=10.68/10.99 Hz, 1H) 6.85 (s, 1H) 6.96 (s, 1H). LC/MS: Anal. Calcd. forC₁₈H₃₄N₃O₃Si [M+H]⁺ 368.24. found 368.43.

Example QC-1e

To a solution of Example QC-1d (2.00 g, 5.45 mmol) in 30 mL of dryacetonitrile at 0° C. was added N-bromosuccinimide (0.97 g, 5.45 mmol)in 30 mL of dry acetonitrile dropwise over 10 min. The resultingsolution was stirred at 0° C. for 2 h. The solvent was removed undervacuum. The residue was taken up in dichloromethane and washed withwater. The aqueous layer was re-extracted with dichloromethane. Theorganic layers were combined, dried with MgSO₄ and concentrated. Thecrude product was purified by flash chromatography (silica gel, 15-40%ethyl acetate/hexanes) to give Example QC-1e as a light yellow oil(1.66, 68.3%). ¹H NMR (500 MHz, CDCl₃) δ ppm −0.02 (s, 9H) 0.79-1.00 (m,2H) 1.21/1.39 (rotomers, s/s, 9H) 1.81-2.08 (m, 2H) 2.10-2.41 (m, 2H)3.36-3.74 (m, 4H) 4.85/4.99 (rotomers, m/m, 1H) 5.26 (m, 1H) 5.39/5.75(rotomers, d/d, J=11.29/11.60 Hz, 1H) 6.93/6.95 (rotomers, s/s, 1H).LC/MS: Anal. Calcd. for C₁₈H₃₃ ⁷⁹BrN₃O₃Si [M]⁺ 446.15. found 446.39.

Example QC-1f

To a solution of Example QC-1b (456 mg, 1.1 mmol), Example QC-1e (982mg, 2.2 mmol), sodium bicarbonate (554 mg, 6.6 mmol) in 21 mL ofdimethylformamide and 7 mL of water was added Pd(PPh₃)₄ (127 mg, 0.11mmol). The resulting solution was refluxed at 85° C. overnight. Thesolvent was removed under vacuum. The residue was taken up in 20%methanol/chloroform and washed with water. The organic layer was washedwith brine, dried with MgSO₄ and concentrated. The crude product waspurified by reserve phase HPLC system (methanol/water/TFA) to giveExample QC-1f as a white solid (560 mg, 57%, mixture of diastereomers).¹H NMR (500 MHz, CDCl₃) δ ppm −0.02 (s, 18H) 0.74-1.03 (m, 4H) 1.19 (s,9H) 1.38 (s, 9H) 1.43-1.59 (m, 2H) 1.76-2.54 (m, 20H) 3.32-3.58 (m, 6H)3.58-3.77 (m, 2H) 4.81 (s, 1H) 4.97 (s, 1H) 5.07-5.21 (m, 2H) 5.21-5.32(m, 1H) 5.60 (t, J=11.29 Hz, 1H) 5.71-5.96 (m, 2H) 6.79 (s, 2H). LC/MS:Anal. Calcd. for C₄₈H₈₁N₆O₆Si₂ [M+H]⁺ 893.58. found 893.42.

Example QC-1 g

To a solution of Example QC-1f (164 mg, 0.18 mmol) in 2 mL ofdichloromethane was added HCl (4 N in 1,4-dioxane, 2 mL). The resultingsolution was stirred at room temperature overnight. The solvent wasremoved under vacuum. The residue was dissolved in minimal methanol andtriturated with diethyl ether. The resulting solid was collected byfiltration and washed with diethyl ether to give the HCl salt of ExampleQC-1 g as a light yellow solid (88.7 mg, 85.2%. mixture ofdiastereomers). LC/MS: Anal. Calcd. for C₂₆H₃₇N₆ [M+H]⁺ 433.31. found433.56.

Example QC-1

To a solution of Example QC-1 g (43.0 mg, 0.074 mmol) and(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (Cap-51) (26.0 mg,0.148 mmol) in 2 mL of dry dimethylformamide was addeddiisopropylethylamine (78 μL, 0.446 mmol). The resulting mixture wasstirred at room temperature for 5 min. HATU (59 mg, 0.156 mmol) was thenadded. The solution was stirred at room temperature for 2 h. The solventwas removed under vacuum. The crude product was purified by reversephase HPLC (methanol/water/TFA). The like fractions were combined,neutralized with saturated NaHCO₃ aqueous solution, and extracted withEtOAc. The organic layers were dried with MgSO₄, and concentrated togive Example QC-1 (free base, mixture of diastereomers) as a white solid(29.5 mg, 53.4%). LC/MS: RT.=2.36 minutes (Cond. 1); LC/MS: Anal. Calcd.for C₄₀H₅₈N₈O₆ [M+H]⁺ 747.46. found 747.72; HRMS: Anal. Calcd. forC₄₀H₅₈N₈O₆ [M+H]⁺ 747.4558. found 747.4587.

Examples QC-2 to QC-5

Examples QC-2 to QC-5 were prepared as free base diasteromeric mixturesby substituting the respective acids for Cap-51 and using the samemethod described for Example QC-1.

Example

RT (LC-Cond); % homogeneity index; MS data QC-2

RT = 2.08 minutes (Cond. 1); LC/MS: Anal. Calcd. for C₃₆H₅₀N₈O₆ [M + H]⁺691.39; found 691.58; HRMS: Anal. Calcd. for C₃₆H₅₀N₈O₆ [M + H]⁺691.3932; found 691.3965 QC-3

RT = 2.10 minutes (Cond. 1); LC/MS: Anal. Calcd. for C₅₀H₆₆N₈O₂ [M + H]⁺811.54; found 811.63; HRMS: Anal. Calcd. for C₅₀H₆₆N₈O₂ [M + H]⁺811.5387; found 811.5426 QC-4

RT = 1.54 minutes (Cond. 2); LC/MS: Anal. Calcd. for C₃₈H₅₄N₈O₈ [M + H]⁺751.41; found 751.34; HRMS: Anal. Calcd. for C₃₈H₅₄N₈O₈ [M + H]⁺751.4143; found 751.4175 QC-5

RT = 1.61 minutes (Cond. 3); LC/MS: Anal. Calcd. for Q₄₀H₅₈N₈O₈ [M + H]⁺779.45; found 779.26

Example QC-6

Example QC-6a

To a solution of Example QC-1 f (554 mg, 0.62 mmol) in 25 mL of methanolin a hydrogenation flask was added palladium (10% on carbon) (125 mg)under nitrogen. The flask was then mixed on the Parr Shaker at 40 atmovernight. After the reaction was complete, the mixture was filtered andthe solvent was removed under vacuum to give Example QC-6a as acolorless glassy diastereomeric mixture (506 mg, 90.9%). LC/MS: Anal.Calcd. for C₄₈H₈₅N₆O₆Si₂ [M+H]⁺ 897.61. found 897.25.

Example QC-6b

To a solution of Example QC-6a (506 mg, 0.56 mmol) in 10 mL ofdichloromethane was added 4N HCl in dioxane (5 mL). The resultingsolution was then stirred at room temperature overnight. The solvent wasremoved under vacuum.

The residue was dissolved in minimal methanol and triturated withdiethyl ether. The resulting solid was collected by filtration andwashed with diethyl ether to give the HCl salt diastereomeric mixture ofExample QC-6b as a light brown solid (300 mg, 92%). LC/MS: Anal. Calcd.for C₂₆H₄₁N₆ [M+H]⁺ 437.34. found 437.41.

Example QC-6

Example QC-6 was synthesized as free base from Example QC-6b and Cap-52according to the procedure described for the synthesis of Example QC-1.LC/MS: RT=2.09 minutes (Cond. 1); LC/MS: Anal. Calcd. for C₃₆H₅₄N₈O₆[M+H]⁺ 695.42. found 695.70; LC/MS: Anal. Calcd. for C₃₆H₅₄N₈O₆ [M+H]⁺695.4245. found 695.4275.

Examples QC-7 and QC-8

Examples QC-7 to QC-8 were prepared as free base diastereomeric mixturesby substituting the respective acids for Cap-52 using the same methoddescribed for Example QC-6.

Example

RT (LC-Cond); % homogeneity index; MS data QC-7

RT = 2.09 minutes (Cond. 1); LC/MS: Anal. Calcd. for C₃₈H₅₈N₈O₈ [M + H]⁺755.45; found 755.73; HRMS: Anal. Calcd. for C₃₈H₅₈N₈O₈ [M + H]⁺755.4456; found 755.4488 QC-8

RT = 1.64 minutes (Cond. 3); LC/MS: Anal. Calcd. for C₄₀H₆₂N₈O₈ [M + H]⁺783.48; found 783.29; HRMS: Anal. Calcd. for C₄₀H₆₂N₈O₈ [M + H]⁺783.4769; found 783.4807

Example QC-9

Example QC-9a

Example QC-9a was synthesized as a diastereomeric mixture from4-(4-hydroxyphenyl)cyclohexanone according to the procedure describedfor the synthesis of Example QC-1a. ¹H NMR (500 MHz, CDCl₃) δ ppm1.89-1.99 (m, 1H) 2.04-2.12 (m, 1H) 2.25-2.61 (m, 4H) 2.85-2.95 (m, 1H)5.83-5.87 (m, 1H) 7.20-7.24 (m, 2H) 7.26-7.31 (m, 2H). LC/MS: Anal.Calcd. for [M+H]⁺ C₁₄H₁₂F₆O₆S₂: 453.99. found (molecule did not ionizewell in mass spec chamber).

Example QC-9b

Example QC-9b was synthesized as a diastereomeric mixture from ExampleQC-9a according to the procedure described for the synthesis of ExampleQC-1b. ¹H NMR (500 MHz, CDCl₃) δ ppm 1.27 (s, 12H) 1.32 (s, 12H)1.62-1.75 (m, 1H) 1.88-1.98 (m, 1H) 2.14-2.46 (m, 4H) 2.73-2.85 (m, 1H)6.60-6.67 (m, J=3.05 Hz, 1H) 7.22 (d, J=8.24 Hz, 2H) 7.74 (d, J=7.93 Hz,2H). LC/MS: Anal. Calcd. for [M+H]⁺ C₂₄H₃₆B₂O₄: 410.28. found (moleculedid not ionize well in mass spec chamber).

Example QC-9c

Example QC-9c was synthesized as a diastereomeric mixture from ExampleQC-1e and Example QC-9b according to the procedure described for thesynthesis of Example QC-1f. ¹H NMR (500 MHz, CDCl₃) δ ppm −0.01 (s, 18H)0.82-1.00 (m, 4H) 1.24 (s, 9H) 1.39 (s, 9H) 1.72-2.68 (m, 14H) 2.73-2.96(m, 1H) 3.29-3.81 (m, 8H) 4.81-5.05 (m, 2H) 5.06-5.23 (m, 2H) 5.31-5.43(m, 1H) 5.76-5.90 (m, 1H) 6.48-6.67 (m, 1H) 6.74 (s, 1H) 7.10 (s, 1H)7.22 (d, J=6.71 Hz, 2H) 7.67 (d, J=7.63 Hz, 2H); LC/MS: Anal. Calcd. forC₄₈H₇₇N₆O₆Si₂ [M+H]⁺ 889.55. found 889.83.

Example QC-9d

Example QC-9d was synthesized from Example QC-9c according to theprocedure described for the synthesis of Example QC-1 g. LC/MS: Anal.Calcd. for C₂₆H₃₃N₆ [M+H]⁺ 429.28. found 429.31.

Example QC-9

Example QC-9 was synthesized as a free base diastereomeric mixture fromExample QC-9d and Cap-51 according to the procedure described for thesynthesis of Example QC-1. LC/MS: RT=2.20 minutes (Cond. 1); LC/MS:Anal. Calcd. for C₄₀H₅₄N₈O₆ [M+H]⁺ 743.42. found 743.31

Examples QC-10 to QC-12

Examples QC-10 to QC-12 were prepared as free base diastereomericmixtures by substituting the respective acids for Cap-51 using the samemethod described for Example QC-9.

Example

RT (LC-Cond); % homogeneity index; MS data QC-10

RT = 1.94 minutes (Cond. 1); LC/MS: Anal. Calcd. for C₃₆H₄₆N₈O₆ [M + H]⁺687.36; found 687.13. QC-11

RT = 1.98 minutes (Cond. 1); LC/MS: Anal. Calcd. for C₃₈H₅₀N₈O₈ [M + H]⁺747.38; found 747.46. QC-12

RT = 2.09 minutes (Cond. 1); LC/MS: Anal. Calcd. for C₄₀H₅₄N₈O₈ [M + H]⁺775.41; found 775.33.

Example QC-13

Example QC-13a

To a solution of 4-iodophenyl ethanone (14.76 g, 60.0 mmol) in 150 mL ofdichloromethane was added bromine (9.5 g, 59.5 mmol) dropwise. Theresulting solution was stirred at room temperature overnight. Thesolvent was removed under vacuum. The crude product was recrystallizedwith dichloromethane/hexanes to give Example QC-13a (11.8 g, 60.5%) as agrey solid. ¹H NMR (500 MHz, CDCl₃) δ ppm 4.38 (s, 2H) 7.68 (d, J=8.55Hz, 2H) 7.86 (d, J=8.85 Hz, 2H); LC/MS: Anal. Calcd. for C₈H₇ ⁷¹BrIO[M+H]⁺ 325.86. found 325.11.

Example QC-13b

To a solution of (S)-1-(benzyloxycarbonyl)pyrrolidine-2-carboxylic acid(6.5 g, 26.1 mmol) in 150 mL of ethyl acetate was added triethylamine(4.0 mL, 28.7 mmol) and Example QC-13a (8.5 g, 26.1 mmol) at 0° C. Theresulting mixture was stirred at room temperature overnight. Thereaction mixture was filtered. The filtrate was washed with saturatedsodium bicarbonate aqueous solution, and brine. The organic layer wasdried with MgSO₄ and concentrated. The crude product was purified by aflash chromatography (silica gel, 10-40% ethyl acetate/hexanes) to givethe product as a white solid Example QC-13b (15.3 g, 85.4%). ¹H NMR (500MHz, CDCl₃) δ ppm 1.87-2.01 (m, 1H) 2.01-2.16 (m, 1H) 2.22-2.42 (m, 2H)3.44-3.58 (m, 1H) 3.59-3.71 (m, 1H) 4.45-4.58 (m, 1H) 5.04-5.50 (m, 4H)7.27-7.38 (m, 5H) 7.53 (d, J=8.24 Hz, 1H) 7.59 (d, J=8.24 Hz, 1H) 7.85(dd, J=8.39, 3.81 Hz, 2H). LC/MS: Anal. Calcd. for C₂₁H₂₁INO₅ [M+H]⁺494.05. found 494.32.

Example QC-13c

A solution of Example QC-13b (15.3 g, 31 mmol) and ammonium acetate(18.0 g, 233 mmol) in 250 mL of xylene was stirred at 110° C. overnight.The reaction mixture was washed with brine, dried with MgSO₄ andconcentrated. The crude product was purified by flash chromatography(silica gel, 25%-65% ethyl acetate/hexanes) to give the product as ayellow solid Example QC-13c (14.1 g, 96.1%). ¹H NMR (300 MHz, CDCl₃) δppm 1.88-2.03 (m, 1H) 2.08-2.33 (m, 2H) 2.82-3.16 (m, 1H) 3.33-3.67 (m,2H) 4.87-5.31 (m, 3H) 7.20 (s, 1H) 7.27-7.41 (m, 5H) 7.42-7.73 (m, 4H)10.38/10.76 (s/s, 1H). LC/MS: Anal. Calcd. for C₂₁H₂₁IN₃O₂ [M+H]⁺474.07. found 474.20.

Example QC-13d

To a solution of Example QC-13c (7.6 g, 16.06 mmol) in 30 mL ofdimethylformamide was added sodium hydride (0.706 g, 17.66 mmol). Theresulting mixture was stirred at room temperature for 40 min.2-(Trimethylsilyl)ethoxymethyl chloride (2.84 ml, 16.06 mmol) was thenadded dropwise over 5 min. After stirred at room temperature for 2 h,the reaction was quenched with water and extracted with ethyl acetate.The combined organic layers were washed with brine, dried with MgSO₄ andconcentrated. The crude product was purified by a flash chromatography(silica gel, 25%-50% ethyl acetate/hexanes) to give Example QC-13d (8.5g, 88%) as an yellow oil [Note: the exact SEM-regiochemical make up ofthe product was not determined] ¹H NMR (500 MHz, CDCl₃) δ ppm−0.02/−0.05 (rotomers, s/s, 9H) 0.73-0.99 (m, 2H) 1.87-2.02 (m, 1H)2.09-2.34 (m, 2H) 2.34-2.48/2.50-2.68 (rotomers, m/m, 1H) 3.26-3.44 (m,1H) 3.45-3.84 (m, 3H) 4.59/4.85 (rotomers, d/d, J=11.29 Hz/12.21 Hz, 1H)4.89-5.13 (m, 3H) 5.16/5.90 (rotomers, d/d, J=10.99 Hz/10.99 Hz, 1H)6.94-7.04 (m, 1H) 7.13-7.37 (m, 5H) 7.50 (d, J=8.55 Hz, 2H) 7.61-7.71(m, 2H). Anal. Calcd. for C₂₇H₃₅IN₃O₃Si [M+H]⁺ 604.15. found 604.44.

Example QC-13e

To a solution of Example QC-1d (8.5 g, 23.13 mmol) in 150 mL of methanolwas added 9 mL of 4 M HCl in dioxane. After stirring at room temperatureovernight, the reaction solution was concentrated under vacuum to give ayellow oil. The yellow oil was placed under vacuum overnight to give(S)-4-iodo-2-(pyrrolidin-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazoleHCl salt as a yellow solid.

To a mixture of(S)-2-(pyrrolidin-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazoleHCl salt (7.03 g, 23.13 mmol) in sodium hydroxide aqueous solution (25.4mL, 50.9 mmol) at 0° C. was added benzyl chloroformate (3.63 mL, 25.4mmol) and sodium hydroxide aqueous solution (2M, 12.72 mL, 25.4 mmol)dropwise at the same time over 1 h. After stirring at 0° C. for 1additional hour, the reaction mixture was extracted with ethyl acetate.The combined organic layers was dried with MgSO₄ and concentrated. Thecrude product was purified by a flash chromatography (silica gel,25%-65% ethyl acetate/hexanes) to give Example QC-13e (8.8 g, 21.9 mmol,95%). ¹H NMR (500 MHz, CDCl₃) δ ppm −0.07/−0.03 (rotomers, s/s, 9H)0.69-0.97 (m, 2H) 1.84-2.00 (m, 1H) 2.05-2.50 (m, 3H) 3.23-3.38 (m, 1H)3.42-3.85 (m, 3H) 4.52/4.87 (rotomers, d/d, J=11.29 Hz, 1H) 4.89-5.14(m, 3H) 5.13/5.85 (rotomers, d/d, J=10.99 Hz, 1H) 6.70/6.87 (rotomers,s/s, 1H) 6.94-6.99 (m, 1H) 6.99-7.05 (m, 1H) 7.25-7.33 (m, 4H). LC/MS:Anal. Calcd. for C₂₁H₃₂N₃O₃Si [M+H]⁺ 402.22. found 402.26.

Example QC-13f

To a solution of Example QC-13e (8.8 g, 21.91 mmol) in 100 mL ofacetonitrile at 0° C. was added NBS (3.90 g, 21.91 mmol) in 60 mL ofacetonitrile dropwise over 10 min. After stirring at 0° C. for 2 h, thesolvent was removed under vacuum. The residue was dissolved indichloromethane and washed with water. The organic layer was dried withMgSO₄ and concentrated. The crude product was purified by a flashchromatography (silica gel, 20%-30% ethyl acetate/hexanes) to giveExample QC-13f (7.5 g, 15.61 mmol, 71.2%) as a yellow oil. ¹H NMR (500MHz, Chloroform-D) δ ppm −0.06/−0.02 (rotomers, s/s, 9H) 0.67-1.00 (m,2H) 1.85-1.99 (m, 1H) 2.03-2.16 (m, 1H) 2.16-2.43 (m, 2H) 3.36 (t,J=8.24 Hz, 1H) 3.48-3.79 (m, 3H) 4.68/4.82 (rotomers, d/d, J=11.60 Hz,1H) 4.85-5.15 (m, 3H) 5.27/5.81 (rotomers, d/d, J=11.29 Hz, 1H)6.93/6.94 (s/s, 1H) 6.95-7.02 (m, 1H) 7.25-7.36 (m, 4H). LC/MS: Anal.Calcd. for C₂₁H₃₀ ⁷⁹BrN₃O₃Si [M]⁺ 479.12. found 479.26.

Example QC-13g

To a mixture of Example QC-13f (4.08 g, 8.49 mmol), tert-butyl4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5,6-dihydropyridine-1(2H)-carboxylate(2.63 g, 8.49 mmol), and sodium bicarbonate (2.140 g, 25.5 mmol) in DME(180 ml) and water (60.0 ml) was addedTetrakis(triphenylphosphine)palladium (0) (0.491 g, 0.425 mmol). Theresulting mixture was stirred at 85° C. overnight. The solvent wasremoved under vacuum. The aqueous mixture was extracted with 20%methanol/chloroform. The organic layer was washed with water, brine,dried with MgSO₄ and concentrated. The crude product was purified by aflash chromatography (silica gel, 25%-50% ethyl acetate/hexanes) to giveExample QC-13g (3.8 g, 6.52 mmol, 76.8%) as a yellow oil. ¹H NMR (500MHz, CDCl₃) δ ppm −0.06/−0.04 (rotomers, s/s, 9H) 0.69-0.97 (m, 2H)1.45/1.47 (rotomers, s/s, 9H) 1.85-1.99 (m, 1H) 2.05-2.28 (m, 2H)2.29-2.63 (m, 3H) 3.20-3.80 (m, 6H) 4.05 (s, 2H) 4.51/4.85 (rotomers,d/d, J=11.29 Hz, 1H) 4.81-5.13 (m, 3H) 5.11/5.86 (rotomers, d/d, J=10.99Hz, 1H) 6.37 (s, 1H) 6.57/6.76 (rotomers, s/s, 1H) 7.01 (d, J=5.49 Hz,1H) 7.19-7.37 (m, 4H). LC/MS: Anal. Calcd. for C₃₁H₄₇N₄O₅Si [M+H]⁺583.33. found 583.65.

Example QC-13h

To a solution Example QC-13g (3.8 g, 6.52 mmol) in 40 mL of methanol wasadded HCl (4N in dioxane, 10 ml, 40.00 mmol). The solution was stirredat room temperature overnight. The solvent was removed under vacuum. Tothe residue was added saturated sodium bicarbonate. The mixture wasextracted with ethyl acetate. The combined organic layers were washedwith brine, dried with MgSO₄ and concentrated. The crude product waspurified by flash chromatography (silica gel, 5%methanol/dichloromethane) to give Example QC-13h (1.90 g, 3.94 mmol,60.4%) as a yellow solid. ¹H NMR (500 MHz, Chloroform-D) δ ppm−0.06/−0.02 (rotomers, s/s, 9H) 0.69-1.05 (m, 2H) 1.78-2.00 (m, 1H)2.01-2.52 (m, 3H) 2.63 (s, 2H) 3.23-3.76 (m, 6H) 3.81 (s, 2H) 4.45-5.87(m, 5H) 6.28 (s, 1H) 6.65/6.84 (rotomers, s/s, 1H) 6.93-7.09 (m, 1H)7.17-7.42 (m, 4H). LC/MS: Anal. Calcd. for C₂₆H₃₉N₄O₃Si [M+H]⁺ 483.28.found 483.55.

Example QC-13i

A mixture of Example QC-13d (3.19 g, 5.28 mmol), Example QC-13h (1.7 g,3.52 mmol), copper(I) iodide (0.134 g, 0.704 mmol), L-proline (0.162 g,1.409 mmol) and potassium carbonate (0.973 g, 7.04 mmol) in DMSO (30 ml)was bubbled with nitrogen for 10 min. The mixture was then stirred at90° C. overnight. Water was added into the reaction. The mixture wasextracted with ethyl acetate. The combined organic layers were washedwith brine, dried with MgSO₄ and concentrated. The crude product waspurified by flash chromatography (silica gel, 10%-40% ethylacetate/hexanes then 40%-75% ethyl acetate/hexanes) to give ExampleQC-13i (1.30 g, 38.6%) as a yellow oil. [Note: the exactSEM-regiochemical make up of the product was not determined] ¹H NMR (300MHz, CDCl₃) δ ppm −0.19-0.10 (m, 18H) 0.65-1.06 (m, 4H) 1.69-2.01 (m,2H) 2.07-2.75 (m, 8H) 3.22-3.99 (m, 12H) 4.60 (t, J=10.61 Hz, 1H)4.77-5.29 (m, 9H) 5.72 (d, J=10.61 Hz, 0.5H) 5.80-5.97 (m, 1H) 6.06 (s,0.5H) 6.84-7.11 (m, 5H) 7.17-7.24 (m, 2H) 7.26-7.39 (m, 6H) 7.62-7.73(m, 2H); LC (Cond. 1): RT=3.28 min; LC/MS: Anal. Calcd. forC₅₃H₇₂N₇O₆Si₂ [M+H]⁺ 958.51. found 958.84.

Example QC-13j

A mixture of Example QC-13i (600 mg, 0.626 mmol) and palladium on carbon(666 mg, 0.626 mmol) in methanol (10 ml) and ethyl acetate (10.00 mL) ina hydrogenation flask and was placed under hydrogen (50 atm.) for 16 h.The catalyst was filtered. To the filtrate was added 6 mL of 4 N HCl in1,4-dioxane, and the resulting solution was stirred at room temperatureovernight. The solvent was removed under vacuum. The residue wastriturated with diethyl ether. The resulting solid was collected byfiltration and washed with diethyl ether to give the HCl salt of ExampleQC-13j (232 mg, 86%) as a brown oil-like solid. ¹H NMR (500 MHz,DMSO-D₆) δ ppm 1.79-2.05 (m, 6H) 2.12-2.23 (m, 2H) 2.29-2.48 (m, 8H)2.80-2.96 (m, 2H) 3.29-3.52 (m, 3H) 4.96-5.10 (m, 2H) 6.93/7.10 (s/s,2H) 7.20-7.49 (m, 4H); LC (Cond. 2): RT=0.69 min; LC/MS: Anal. Calcd.for C₂₅H₃₄N₇ [M+H]⁺ 432.29. found 432.30.

Example QC-13

Example QC-13 was synthesized from Example QC-14j and Cap-51 accordingto the procedure described for the synthesis of Example QC-1, with theexception that the final purified material was not neutralized andExample QC13 was obtained as TFA salt. LC/MS: RT=1.95 min. (Cond. 1);LC/MS: Anal. Calcd. for C₃₉H₅₆N₉O₆ [M+H]⁺ 746.44. found 746.97.

Examples QC-14 and QC-15

Examples QC-14 to QC-15 were prepared as TFA salts by substituting therespective acids for Cap-51 using the same method described for ExampleQC-13.

Example

RT (LC-Cond); % homogeneity index; MS data QC-14

RT = 1.64 minutes (Cond. 1); LC/MS: Anal. Calcd. for C₃₇H₅₂N₉O₈ [M + H]⁺750.39; found 750.92; QC-15

RT = 1.93 minutes (Cond. 1); LC/MS: Anal. Calcd. for C₄₁H₆₀N₉O₆ [M + H]⁺774.47; found 774.76;

Example QC-16 and QC-17

Example QC-16a

To a solution of Example QC-9c (260 mg, 0.3 mmol) in 10 mL of methanolwas added palladium (10% on carbon) (100 mg) under nitrogen. The flaskwas then vacuumed and flushed with N₂ for 3 times. After being vacuumedagain, the mixture was hydrogenated under H₂ balloon overnight. Afterthe reaction was complete, the mixture was filtered and the solvent wasremoved under vacuum to give Example QC-16a as a greasy soliddiastereomeric mixture. LC/MS: RT=3.28 minutes (Cond. 1); LC/MS: Anal.Calcd. for C₄₈H₈₇N₆O₆Si₂ [M+H]⁺ 891.56. found 891.64.

Example QC-16b

Example QC-16b was synthesized as a diastereomeic mixture from ExampleQC-16a according to the procedure described for the synthesis of ExampleQC-1g. LC/MS: RT=1.53 minutes and 1.64 minutes (Cond. 1); LC/MS: Anal.Calcd. for C₂₆H₃₅N₆ [M+H]⁺ 431.29. found 431.25 and 431.26.

Example QC-16 and QC-17

Example QC-16 and QC-17 were synthesized as free bases from ExampleQC-13b and Cap-51 according to the procedure described for the synthesisof Example QC-1. The crude residue was purified by reverse phase HPLC(Acetonitrile/Water/NH₄OAc) to give two diastereomers QC-16 and QC-17.QC-16: ¹HNMR (300 MHz, DMSO-D₆) δ ppm 0.73-0.93 (m, 12H), 1.45-2.30 (m,18H), 2.40-2.65 (m, overlap with DMSO solvent peak), 3.15-3.45 (m,overlap with water peak), 3.52 (s, 6H), 3.75 (m, 3H), 4.01 (m, 2H), 5.03(m, 2H), 7.00-7.70 (m, 6H), 11.23 (d, 1H), 11.66 (br, 1H); LC/MS:RT=2.20 minutes (Cond. 1); LC/MS: Anal. Calcd. for C₄₀H₅₆N₈O₆ [M+H]⁺745.44. found 745.31; QC-17: ¹H NMR (500 MHz, DMSO-D₆) δ ppm 0.75-0.93(m, 12H), 1.30-1.62 (m, 4H), 1.75-2.22 (m, 14H), 2.40-2.65 (m, overlapwith DMSO solvent peak), 3.15-3.45 (m, overlap with water peak), 3.53(s, 6H), 3.78 (m, 3H), 4.04 (m, 2H), 5.03 (m, 2H), 7.10-7.70 (m, 6H),11.25 (d, 1H), 11.68 (br, 1H); LC/MS: RT=2.20 minutes (Cond. 1); LC/MS:Anal. Calcd. for C₄₀H₅₆N₈O₆ [M+H]⁺ 745.44. found 745.37.

Example M-1

Example M-1a

A dichloromethane (13 mL) solution of bromine (2.25 g, 14.1 mmol) wasadded drop-wise over 10 min to a heterogeneous refluxing mixture of4-(methoxycarbonyl)bicycle [2.2.2]octane-1-carboxylic acid (2.02 g, 9.52mmol) and mercuric oxide (3.5 g, 16.16 mmol) in dichloromethane (40 mL),and heating was continued for 3.3 h. After the reaction mixture wasallowed to cool to room temperature, it was filtered and the resultinglight orange filtrate was treated with MgSO₄ and filtered again. Thevolatile component of the filtrate was removed in vacuo, and theresulting residue was purified with a BIOTAGE® (5-10% ethylacetate/hexanes) to afford Example M-1a as a white solid (1.37 g). ¹HNMR (DMSO-d₆, δ=2.50, 400 MHz): 3.56 (s, 3H), 2.22-2.18 (m, 6H),1.91-1.87 (m, 6H). LC (Cond. 2): RT=1.79 min. LC/MS: Anal. Calcd. for[M+H]⁺ C₁₀H₁₆BrO₂: 247.03. found 247.33.

Example M-1b

A benzene (65 mL) solution of Example M-1a (3.758 g, 15.21 mmol) wasadded drop-wise to a cooled (˜−12° C.) mixture of benzene (200 mL) andaluminum trichloride (8.93 g, 67 mmol) over 15 min. The heterogeneousmixture was stirred for 1 h while allowing the cooling bath to thawgradually to ˜3° C., and then the cooling bath was removed and stirringwas continued for ˜14 h. The reaction mixture was then heated for ˜4 hr,and after the mixture was allowed to cool to ambient condition and mostof the volatile component was removed in vacuo. The residue was taken updichloromethane (100 mL), poured into 150 mL of ice-water, and thephases were separated. The aqueous phase was washed with dichloromethane(50 mL), and the combined organic phase was washed with saturated sodiumbicarbonate, dried (MgSO₄) and evaporated in vacuo. A silica gel meshwas prepared from the resulting crude material and submitted to flashchromatography (3-4.5% ethyl acetate/hexanes) to afford Example M-1b asan off-white solid (2.739 g). ¹H NMR (DMSO-d₆, δ=2.50, 400 MHz):7.34-7.26 (m, 4H), 7.18-7.13 (m, 1H), 3.59 (s, 3H), 1.86-1.76 (m, 12H).LC (Cond. 2): RT=2.04 min. LC/MS: Anal. Calcd. for [M+H]⁺ C₁₆H₂₁O₂:245.15. found 245.24.

Example M-1c

A chloroform (16 mL) solution of bromine (2.07 g, 12.95 mmol) was addeddrop-wise, via an addition funnel, over 13 min to a vigorously stirredmixture of Example M-1b (3.01 g, 12.33 mmol) and silver trifluoroacetate(3.15 g, 14.26 mmol) in chloroform (45 mL). After stirring for 105 min,the precipitate was filtered and washed with chloroform, and thefiltrate was evaporated in vacuo. The resultant crude material waspurified with flash chromatography (sample was loaded as a silica gelmesh; 4-6% ethyl acetate/hexanes) to afford Example M-1c as a whitesolid (3.495 g). ¹H NMR (DMSO-d₆, δ=2.50, 400 MHz): 7.46 (d, J=8.5, 2H),7.28 (d, J=8.8, 2H), 3.59 (s, 3H), 1.82-1.76 (m, 12H). LC (Cond. 2):RT=2.16 min. LC/MS: Anal. Calcd. for [M+H]⁺ C₁₆H₂₀BrO₂: 323.06. found323.18.

Example M-1d

Butyl lithium (1.6 M/hexanes) (19.5 mL, 31.2 mmol) was added drop-wiseover 15 min to a cooled (ice-water) tetrahydrofuran (10 mL) solution ofi-Pr₂NH (4.5 mL, 31.6 mmol) and stirred for 25 min. The resultant LDAsolution was added drop-wise over 15 min to a cooled (−78° C.)tetrahydrofuran (30 mL) solution of Example M-1c (2.48 g, 7.68 mmol) andchloroiodomethane (5.57 g, 31.6 mmol), and stirred for 130 min. Atetrahydrofuran (30 mL) solution of acetic acid (12 mL) was addeddrop-wise over 10 min to the above reaction mixture, and 20 min laterthe cooling bath was removed and stirring was continued for 30 min. Thenthe volatile component was removed in vacuo, and the residue was takenup in ethyl acetate (100 mL) and washed with saturated sodiumbicarbonate solution (20 ml, 4×) and sodium thiosulfate solution (3.3 gin 30 mL water), dried (MgSO₄) and evaporated in vacuo. A silica gelmesh was prepared from the resulting crude material and submitted toflash chromatography (30-50% dichloromethane/hexanes) to afford ExampleM-1d as an off-white solid (2.02 g). ¹H NMR (DMSO-d₆, δ=2.50, 400 MHz):7.46 (d, J=8.6, 2H), 7.29 (d, J=8.6, 2H), 4.78 (s, 2H), 1.82-1.76 (m,12H). HRMS: Anal. Calcd. for [M] C₁₆H₁₈BrClO: 340.0230. found 340.0234.

Example M-1e

Diformylimide sodium salt (0.7187 g, 7.18 mmol) and potassium iodide(0.148 g, 0.892 mmol) were added to a tetrahydrofuran (12.5 mL) solutionof Example M-1d (1.01 g, 2.96 mmol), and the heterogeneous mixture wassonicated for a few minutes and then stirred at ambient condition for˜23 h. The reaction mixture was filtered, the filtered solid was washedwith dichloromethane, and the filtrate was evaporated in vacuo. Theresulting crude solid was transferred to a 350 mL pressure tubecontaining methanol (40 mL), water (20 mL) and concentrated HCl (2.5mL), and the heterogeneous mixture was heated at 60° C. for 23 h.Removal of the volatile component in vacuo afforded Example M-1e as alight yellow solid (1.078 g), which was used in the next step withoutpurification. ¹H NMR (DMSO-d₆, δ=2.50, 400 MHz): 8.01 (br s, ˜3H), 7.48(d, J=8.6, 2H), 7.30 (d, J=8.6, 2H), 4.06 (s, 2H), 1.80 (app s, 12H). LC(Cond. 2): RT=1.58 min. LC/MS: Anal. Calcd. for [M+H]⁺ C₁₆H₂₁BrNO:322.08. found 322.21.

Example M-1f

i-Pr₂EtN (1.02 mL, 2.5 mmol) was added drop-wise over a few minutes to amixture of Example M-1e (0.84 g, 2.35 mmol), Boc-L-Proline (0.657 g,3.05 mmol) and HATU (1.07 g, 2.82 mmol) in dimethylformamide (15 mL),and the reaction mixture was stirred for 50 min. The volatile componentwas removed in vacuo, and the residue was taken up in dichloromethane(80 mL) and washed with water (25 mL, 2×) and saturated sodiumbicarbonate (25 mL, 2×). The organic layer was dried (MgSO₄) andevaporated in vacuo. The resultant crude material was submitted to aBIOTAGE® purification (100 g silica gel; 40-70% ethyl acetate/hexanes)to afford Example M-1f as a white dense solid (938 mg); a slightlyimpure sample of Example M-1f was also retrieved (118 mg). ¹H NMR(DMSO-d₆, δ=2.50, 400 MHz): 7.92-7.85 (m, 1H), 7.47 (d, J=8.8, 2H), 7.29(d, J=8.8, 2H), 4.16-4.01 (m, 3H), 3.40-3.24 (m, 2H; partiallyoverlapped with water signal), 2.15-2.00 (m, 1H), 1.89-1.70 (m, 15H),1.39/1.33 (two overlapped s, 9H). LC (Cond. 2): RT=2.10 min. LC/MS:Anal. Calcd. for [M+H]⁺ C₂₆H₃₆BrN₂O₄: 519.19. found 519.18.

Example M-1g

A mixture of Example M-1f (1.044 g, 2.01 mmol) and ammonium acetate(1.260 g, 16.35 mmol) in xylenes (12 mL) was heated with a microwave at140° C. for 2 h. An additional ammonium acetate (240 mg, 3.11 mmol) wasadded and the mixture heated similarly for 30 min. Then the volatilecomponent was removed in vacuo, and the residue was treated withdichloromethane (80 mL), water (20 mL) and saturated sodium bicarbonatesolution (4 mL), and vigorously stirred and the phases were separated.The organic layer was dried (MgSO₄) and evaporated in vacuo. The crudematerial was purified with a BIOTAGE® (60-100% ethyl acetate/hexanes) toretrieve Example M-1g as a white foam (716 mg). ¹H NMR (MeOH-d₄, δ=3.29,400 MHz): 7.39 (d, J=8.6, 2H), 7.26 (d, J=8.8, 2H), 6.57/6.54(overlapping br s, 1H), 4.78-4.72 (m, 1H), 3.66-3.57 (m, 1H), 3.48-3.41(m, 1H), 2.32-1.86 (overlap of br m & app s, 16H), 1.43 (app s, 2.61H),1.20 (s, 6.39H). LC (Cond. 2): RT=1.80 min. LC/MS: Anal. Calcd. for[M+H]⁺ C₂₆H₃₅BrN₃O₂: 500.19. found 500.11.

Example M-1 h

Sodium hydride (60%; 57 mg, 1.43 mmol) was added in one batch to adimethylformamide (6 mL) solution of the imidazole Example M-1g (503 mg,1.005 mmol), and the mixture was stirred for 70 min. Then, SEM-Cl (0.21mL, 1.187 mmol) was added drop-wise over 30 sec to the above reactionmixture, and the heterogeneous mixture was stirred for 4.3 h. Thevolatile component was removed in vacuo, and the residue was partitionedbetween dichloromethane and water. The organic layer was dried (MgSO₄)and evaporated in vacuo. The resulting crude material was purified witha BIOTAGE® (20-35% ethyl acetate/hexanes) to afford Example M-1 h as awhite foam (417 mg). ¹H NMR (DMSO-d₆, δ=2.50, 400 MHz): 7.46 (d, J=8.6,2H), 7.31 (d, J=8.8, 2H), 6.78/6.75 (two overlapping s, 1H), 5.54 (br d,J=10.5, 0.32H), 5.28 (br d, J=11.1, 0.68H), 5.17 (app br d, J=10.8, 1H),4.88 (br m, 0.32H), 4.78 (br m, 0.68H), 3.5-3.3 (m, 4H), 2.28-1.73(overlap of br m & app s, 16H), 1.35 (s, 2.9H), 1.11 (s, 6.1H),0.89-0.75 (m, 2H), −0.04 (s, 9H). LC/MS (Cond. 2): Anal. Calcd. for[M+H]⁺ C₃₂H₄₉BrN₃O₃Si: 630.27. found 630.33.

Example M-1i

Palladium(II) acetate (15.7 mg, 0.070 mmol) was added in one batch to a75 mL pressure tube containing Example M-1 h (539 mg, 0.855 mmol),Example QC-1d (409.3 mg, 1.14 mmol), triphenylphosphine (35.5 mg, 0.135mmol) and potassium carbonate (132 mg, 0.958 mmol) in dioxane (8 mL),and the reaction mixture was flushed with nitrogen for 5 min and heatedwith 120° C. oil bath for 16 h. After it was allowed to cool to ambientcondition, the mixture was filtered and the filtrate was rotervaped. Theresulting crude material was purified with a combination of BIOTAGE®(100 g silica gel; 40-80% ethyl acetate/hexanes) and a reverse phaseHPLC (water/methanol/TFA). The combined HPLC fractions was neutralizedwith excess NH₃/methanol, rotervaped, and the resulting material waspartitioned between dichloromethane (50 mL), water (30 mL) and satd.sodium bicarbonate solution (1 mL). The organic phase was dried (MgSO₄)and evaporated in vacuo to afford Example M-1i as a white foam (352 mg).The exact SEM regiochemical make up was not determined as it wasinconsequential for the current purpose. ¹H NMR (DMSO-D₆, δ=2.50, 400MHz): 7.45-7.35 (m, 4H), 6.93/6.91 (overlapping s, 1H), 6.79/6.76(overlapping s, 1H), 5.56-5.52 (m, 0.7H), 5.37-5.16 (m, 3.3H), 5.05-4.78(m, 2H), 3.56-3.33 (m, 8H), 2.29-1.76 (m, 20H), 1.37 (s, 6.53),1.13/1.12 (overlapping s, 11.47H), 0.90-0.76 (m, 4H), −0.03/−0.06/−0.07(overlapping s, 18H). LC/MS (Cond. 2): Anal. Calcd. for [M-SEM+H]⁺C₄₄H₄₄N₆O₅Si: 786.49. found 787.50.

Example M-1 j

A mixture of water (2 mL) and concentrated HCl (1 mL) was added to amixture of HCl/dioxanes (4.0 N, 7 mL) and Example M-1i (348 mg, 0.379mmol), and the resultant solution was stirred at ambient condition untilLC/MS analysis indicated completion of reaction (˜48 h). The volatilecomponent was removed in vacuo to afford Example M-1j (0.4HCl) as anoff-white solid, weighing 244.6 mg (˜16 mg above the theoretical yield).The sample contained unidentified impurities, and was used in the nextstep with out purification. LC (Cond. 2): RT=0.86 min. LC/MS: Anal.Calcd. for [M+H]^(+C) ₂₈H₃₇N₆: 457.31. found 457.36.

Example M1

HATU (106.2 mg, 0.279 mmol) was added to a dimethylformamide (2 mL)solution of Example M-1j (4HCl salt) (78 mg, 0.129 mmol),(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (57.7 mg, 0.329 mmol)and i-Pr₂EtN (0.15 mL, 0.859 mmol), and the reaction mixture was stirredfor 30 min. The volatile component was removed in vacuo, and the crudematerial was first passed through MCX (2 g; methanol wash; 2.0 Mammonia/methanol elution) and then submitted to a reverse phase HPLCpurification (water/methanol/TFA) to afford a hygroscopic material. Thepurified material was free-based (MCX; methanol wash; 2.0 Mammonia/methanol elution) and dried in vacuo to afford Example M-1 as anoff-white foam (43 mg). ¹H NMR (DMSO-D₆, δ=2.50, 400 MHz): 12.16-11.18(collections of s, ˜2H), 7.65-6.37 (m, 8H), 5.30-4.99 (m, 2H), 4.06-3.99(m, 2H), 3.83-3.65 & 3.55-3.49 (m, 4H), 3.525/3.53/3.41 (three s, 6H),2.18-1.75 (m, 22H), 0.89-0.81 (m, 12H). LC (Cond. 2): RT=1.33 min.LC/MS: Anal. Calcd. for [M+H]⁺ C₄₂H₅₉H₈O₆: 771.46. found 771.47.

Examples M-2 and M-3

Examples M2 and M3 were prepared as TFA salts from Example M-1j andappropriate acids by employing the procedure described for the synthesisof Example M-1, with the exception that an additional purification wasconducted with a second reverse phase HPLC system(water/acetonitrile/TFA) and that the final purified material was notfree-based.

Example

RT (LC-Cond.); % homogeneity index; MS data M-2

1.13 min (Cond. 2); >95%; LC/MS: Anal. Calcd. for [M + H]⁺ C₃₈H₅₁N₈O₆:715.39; found 715.45 M-3

1.36 min (Cond. 2); >95%; LC/MS: Anal. Calcd. for [M + H]⁺ C₄₈H₅₅N₈O₆:839.42; found 839.32

Example M-4

Example M-4-a

Hunig's base (500 μL, 2.87 mmol) was added drop-wise over ˜1 min to aacetonitrile (5 mL)/dichloromethane (5 mL) semi-solution of(S)-1-(tert-butoxycarbonyl)pyrrolidine-2-carboxylic acid (645 mg, 3.00mmol), Example M-1d (405 mg, 1.185 mmol) and potassium iodide (60.5 mg,0.364 mmol), and the reaction mixture was stirred at ambient conditionfor ˜18 h. The volatile component was removed in vacuo and the residuewas partitioned between dichloromethane and water, dried (MgSO₄) andevaporated in vacuo. The resulting crude material was submitted to aBIOTAGE® purification (15-30% ethyl acetate/hexanes) to afford ExampleM-4-a as a white foam (559 mg). ¹H NMR (DMSO-D₆, δ=2.50, 400 MHz): 7.47(d, J=8.5, 2H), 7.29 (d, J=8.8, 2H), 5.13-4.91 (m, 2H), 4.29-4.23 (m,1H), 3.39-3.26 (m, 2H), 2.30-2.16 (m, 1H), 2.11-2.03 (m, 1H), 1.89-1.73(m, 14H), 1.39 (s, 3.59H), 1.34 (s, 5.41H). LC (Cond. 2): RT=2.20 min.LC/MS: Anal. Calcd. for [M-Boc+H]⁺ C₂₁H₂₇BrNO₃: 420.12. found 420.13.

Example M-4-b

Ammonium acetate (825 mg, 10.70 mmol) was added into a toluene (10 mL)solution of Example M-4-a (556 mg, 1.068 mmol), and the mixture washeated under a Dean-Stark set up with a ˜120° C. oil bath for ˜5.7 h.After it was allowed to cool to ambient condition, the volatilecomponent was removed in vacuo, and the residue was carefullypartitioned between dichloromethane (50 mL) and 50% saturated sodiumbicarbonate solution (20 mL). The organic layer was dried (MgSO₄), andevaporated in vacuo. The resulting crude material was purified with aBIOTAGE® (0-20% ethyl acetate/dichloromethane) to afford Example M-4-b(430 mg) as a dense solid, and Example M-1g (12 mg) as a white film.Example M-4-b: ¹H NMR (DMSO-D₆, δ=2.50, 400 MHz): 7.67/7.63 (overlappeds, 1H), 7.47 (d, J=8.5, 2H), 7.31 (d, J=8.8, 2H), 4.85-4.73 (m, 1H),3.48-3.42 (m, 1H), 3.38-3.32 (m, 1H), 2.29-2.15 (m, 1H), 1.95-1.75(15H), 1.38 (s, 2.21H), 1.18 (s, 6.79H). LC (Cond. 2): RT=2.28 min.LC/MS: Anal. Calcd. for [M+H]⁺ C₂₆H₃₄BrN₂O₃: 501.18. found 501.15.

Example M-4-c

Palladium (II) acetate (11.1 mg, 0.049 mmol) was added in one batch to a75 mL pressure tube containing Example M-4-b (412 mg, 0.822 mmol),Example QC-1d (323 mg, 0.879 mmol), triphenylphosphine (25.1 mg, 0.096mmol) and potassium carbonate (139 mg, 1.004 mmol) in dioxane (7 mL),and the reaction mixture was flushed with nitrogen for a few minutes andheated with 122° C. oil bath for ˜17 h. After it was allowed to cool toambient condition, the mixture was filtered, and the filtrate wasrotervaped. The resulting crude material was purified with a combinationof BIOTAGE® (100 g silica gel; 40-100% ethyl acetate/hexanes) andreverse phase HPLC (water/methanol/TFA). The combined HPLC fraction wasneutralized with excess ammonia/methanol, concentrated in vacuo, and theresulting material was partitioned between dichloromethane (50 mL) anddilute aqueous sodium bicarbonate solution (˜6%, 32 mL). The organicphase was dried (MgSO₄) and evaporated in vacuo to afford the coupledproduct Example M-4-c as a white foam (162 mg). The exact regioisomericmake up of the SEM protecting group was not determined as it isinconsequential for the current purpose. ¹H NMR (DMSO-D₆, δ=2.50, 400MHz): 7.67/7.64 (overlapped s, 1H), 7.45-7.35 (m, 4H), 6.93/6.91(overlapped s, 1H), 5.54-4.74 (m, 4H), 3.57-3.33 (m, 6H), 2.29-1.77 (m,20H), 1.38/1.37 (overlapped s, 5.85H), 1.19/1.13 (overlapped s, 12.15H),0.83-0.76 (2H), −0.06/−0.07 (overlapped s, 9H). LC (Cond. 2): RT=2.09min. LC/MS: Anal. Calcd. for [M+H]⁺ C₄₄H₆₆N₅O₆Si: 788.48. found 788.54.

Example M-4-d

HCl (6 mL of 4.0 N HCl/dioxane) was added to Example M-4-c (158.1 mg,0.201 mmol) and the mixture was stirred at ambient condition for 15 h,resulting in a formation of a heavy suspension. An aqueous HCl solution(1 mL of a solution prepared from 2 mL water and 1 mL concentrated HCl)was added to the above mixture and stirring was continued for 23.5 h.Removal of the volatile component in vacuo afforded the HCl salt ofExample M-4-d, containing unidentified impurity, as off-white fluffysolid (118 mg). LC (Cond. 2): RT=1.06 min. LC/MS: Anal. Calcd. for[M+H]⁺ C₂₈H₃₆N₅O: 458.29. found 458.32.

Example M-4

Example M-4 (TFA salt) was prepared from Example M-4-d according to theprocedure described for the synthesis of Example M-1. ¹H NMR (DMSO-D₆,δ=2.50, 400 MHz): 14.50 (app br s, 2H), 8.02 (s, 1H), 7.67 (d, J=8.3,2H), 7.63 (s, 1H), 7.52 (d, J=8.6, 2H), 7.35 (d, J=8.3, 2H), 5.13-5.09(m, 1H), 5.03 (dd, J=3.9, 1H), 4.12-4.08 (m, 1H), 4.06-4.02 (m, 1H),3.89-3.70 (m, 4H), 3.54/3.52 (overlapping s, 6H), 2.43-2.36 (m, 1H),2.23-1.77 (m, 21H), 0.91-0.76 (12H). [Note: a minor component presumedto be a rotamer appears to be present and its chemical shift was notincluded]. LC (Cond. 2): RT=1.68 min. LC/MS: Anal. Calcd. for [M+H]⁺C₄₂H₅₈N₇O₇: 772.44. found 772.51.

Example M-5

Example M-5a

To a solution of (S)-5-(hydroxymethyl)pyrrolidin-2-one (10 g, 87 mmol)in dichloromethane (50 mL) was added tert-butylchlorodiphenylsilane(25.6 g, 93 mmol), triethylamine (12.1 mL, 87 mmol) and DMAP (1.06 g,8.7 mmol). The mixture was stirred at room temperature until thestarting pyrrolidinone was completely consumed, and then it was dilutedwith dichloromethane (50 mL) and washed with water (50 mL). The organiclayer was dried (Na₂SO₄), filtered, and evaporated in vacuo, and thecrude material was submitted to flash chromatography (silica gel; 30 to100% of ethyl acetate/hexanes) to afford the silyl ether as a colorlessoil (22.7 g, 74% yield). ¹H NMR (400 MHz, DMSO-d₆, δ=2.5 ppm) 7.69 (brs, 1H), 7.64-7.61 (m, 4H), 7.50-7.42 (m, 6H), 3.67-3.62 (m, 1H),3.58-3.51 (m, 2H), 2.24-2.04 (m, 3H), 1.87-1.81 (m, 1H), 1.00 (s, 9H).

Di-tert-butyl dicarbonate (38.5 g, 177 mmol) was added in portions as asolid over 10 min to a dichloromethane (200 mL) solution of the silylether prepared above (31.2 g, 88.3 mmol), triethylamine (8.93 g, 88mmol), and DMAP (1.08 g, 8.83 mmol) and stirred for 18 h at 24° C. Mostof the volatile material was removed in vacuo and the crude materialtaken up in 20% ethyl acetate/hexanes and applied to a 2 L funnelcontaining 1.3 L of silica gel and then eluted with 3 L of 20% ethylacetate/hexane and 2 L of 50% ethyl acetate). Upon concentration of thedesired fractions in a rotary evaporator, a white slurry of solid formedwhich was filtered, washed with hexanes and dried in vacuo to affordcarbamate Example M-5a as a white solid (32.65 g, 82% yield). ¹H NMR(400 MHz, DMSO-d₆, δ=2.5 ppm) 7.61-7.59 (m, 2H), 7.56-7.54 (m, 2H),7.50-7.38 (m, 6H), 4.18 (m, 1H), 3.90 (dd, J=10.4, 3.6, 1H), 3.68 (dd,J=10.4, 2.1, 1H), 2.68-2.58 (m, 1H), 2.40-2.33 (m, 1H), 2.22-2.12 (m,1H), 2.01-1.96 (m, 1H), 1.35 (s, 9H), 0.97 (s, 9H). LC (Cond. 2):RT=2.18 min. LC/MS: Anal. Calcd. for [M+Na]⁺ C₂₆H₃₅NNaO₄Si: 476.22.found 476.14.

Example M-5b

A three-necked flask equipped with a thermometer and a nitrogen inletwas charged with Example M-5a (10.05 g, 22.16 mmol) and toluene (36 mL),and lowered into −55° C. cooling bath. When the internal temperature ofthe mixture reached −50° C., lithium triethylborohydride (23 mL of 1.0M/tetrahydrofuran, 23 mmol) was added dropwise over 30 min and themixture stirred for 35 min while maintaining the internal temperaturebetween −50° C. and −45° C. Hunig's base (16.5 mL, 94 mmol) was addeddropwise over 10 min. Then, DMAP (34 mg, 0.278 mmol) was added in onebatch, followed by the addition of trifluoroacetic anhydride (3.6 mL,25.5 mmol) over 15 min, while maintaining the internal temperaturebetween −50° C. and −45° C. The cooling bath was removed 10 min later,and the reaction mixture was stirred for 14 h while allowing it to riseto ambient temperature. It was diluted with toluene (15 mL), cooled withan ice-water bath, and treated slowly with water (55 mL) over 5 min. Thephases were separated and the organic layer washed with water (50 mL,2×) and concentrated in vacuo. The crude material was purified by flashchromatography (silica gel; 5% ethyl acetate/hexanes) to afford ExampleM-5b as a colorless viscous oil (7.947 g, 82% yield). ¹H NMR (400 MHz,DMSO-d₆, δ=2.5 ppm) 7.62-7.58 (m, 4H), 7.49-7.40 (m, 6H), 6.47 (br s,1H), 5.07/5.01 (overlapping br d, 1H), 4.18 (br s, 1H), 3.89 (br s,0.49H), 3.69 (br s, 1.51H), 2.90-2.58 (br m, 2H), 1.40/1.26 (overlappingbr s, 9H), 0.98 (s, 9H). LC (Cond. 2): RT=2.41 min. LC/MS: Anal. Calcd.for [M+Na]⁺ C₂₆H₃₅NNaO₃Si: 460.23. found 460.19.

Examples M-5c-1 and M-5c-2

Diethylzinc (19 mL of ˜1.1 M in toluene, 20.9 mmol) was added dropwiseover 15 min to a cooled (−30° C.) toluene (27 mL) solution ofdihydropyrrole Example M-5b (3.94 g, 9.0 mmol). Chloroiodomethane(stabilized over copper; 3.0 mL, 41.2 mmol) was added dropwise over 10min, and stirred while maintaining the bath temperature at −25° C. for 1h and between −25° C. and −21° C. for 18.5 h. The reaction mixture wasopened to the air and quenched by the slow addition of 50% saturatedsodium bicarbonate solution (40 mL), and then removed from the coolingbath and stirred at ambient temperature for 20 min. It was filteredthrough a filter paper and the white cake was washed with 50 mL oftoluene. The organic phase of the filtrate was separated and washed withwater (40 mL, 2×), dried (MgSO₄) and concentrated in vacuo. The crudematerial was purified using a BIOTAGE® system (350 g silica gel; samplewas loaded with 7% ethyl acetate/hexanes; eluted with 7-20% ethylacetate/hexanes) to afford Examples M-5c-1/5c-2 as a colorless viscousoil, mainly as the trans isomer (3.691 g, 90.7%). [Note: the exacttrans/cis ratio was not determined at this stage]. ¹H NMR (DMSO-d₆,δ=2.50, 400 MHz) of Example M-5c-1: 7.62-7.60 (m, 4H), 7.49-7.40 (m,6H), 3.76 (br m, 1H), 3.67 (br m, 2H), 3.11-3.07 (m, 1H), 2.23 (br m,1H), 2.03 (br m, 1H), 1.56-1.50 (m, 1H), 1.33 (br s, 9H), 1.00 (s, 9H),0.80-0.75 (m, 1H), 0.30 (br m, 1H). LC (Cond. 2): RT=2.39 min. LC/MS:Anal. Calcd. for [M+Na]⁺ C₂₇H₃₇NNaO₃Si: 474.24. found 474.14.

Examples M-5d-1 and M-5d-2

TBAF (7.27 mL of 1.0 M in THF, 7.27 mmol) was added dropwise over 5 minto a THF (30 mL) solution of Examples M-5c-1/−5c-2 (3.13 g, 6.93 mmol)and the mixture was stirred at ambient condition for 4.75 hours. Afterit was treated with saturated NH₄Cl solution (5 mL), most of thevolatile component was removed in vacuo and the residue was partitionedbetween CH₂Cl₂ (70 mL) and 50% saturated NH₄Cl solution (30 mL). Theaqueous phase was extracted with CH₂Cl₂ (30 mL), and the combinedorganic phase was dried (MgSO₄), filtered, concentrated in vacuo, andthen exposed to high vacuum overnight. The resulting crude material waspurified with a BIOTAGE® (40-50% EtOAc/hexanes) to afford ExamplesM-5d-1/5d-2 as colorless oil, mainly as the trans isomer, contaminatedwith traces of lower R_(f) impurities (1.39 g, ˜94%). [Note: the exacttrans/cis ratio was not been determined at this stage]. ¹H NMR (DMSO-d₆,δ=2.50, 400 MHz) of Example M-5d-1: 4.70 (app t, J=5.7, 1H), 3.62-3.56(m, 1H), 3.49-3.44 (m, 1H), 3.33-3.27 (m, 1H), 3.08-3.04 (m, 1H), 2.07(br m, 1H), 1.93-1.87 (m, 1H), 1.51-144 (m, 1H), 1.40 (s, 9H), 0.76-0.71(m, 1H), 0.26 (br m, 1H).

Example M-5e

A semi-solution of NaIO₄ (6.46 g, 30.2 mmol) in H₂O (31 mL) was added toa solution of Examples M-5d-1/−5d-2 (2.15 g, 10.08 mmol) in CH₃CN (20mL) and CCl₄ (20 mL). R^(u)Cl₃ (0.044 g, 0.212 mmol) was addedimmediately and the heterogeneous reaction mixture was vigorouslystirred for 75 minutes. The reaction mixture was diluted H₂O (60 mL) andextracted with CH₂Cl₂ (50 mL, 3×). The combined organic phase wastreated with 1 mL CH₃OH, allowed to stand for about 5 min, and thenfiltered through a pad of diatomaceous earth (CELITE®). The CELITE® waswashed with CH₂Cl₂ (50 mL), and the filtrate was rotervaped to afford alight charcoal-colored solid. ¹H NMR of the crude material indicated a1.00:0.04:0.18 mole ratio among trans acid M-5e-1:presumed cis acidM-5e-2:side product M-5e-3. The crude material was dissolved in EtOAc(˜10 mL) with heating, and allowed to stand at ambient condition withseeding. About 15 minutes into the cooling phase, a rapid crystalformation was observed. About 1 hour later, hexanes (˜6 mL) was addedand the mixture was refrigerated overnight (it did not appear thatadditional compound has precipitated out). The mixture was filtered andwashed with ice/water cooled hexanes/EtOAc (2:1 ratio; 20 mL) and driedunder high vacuum to afford the first crop of Example M-5e-1 (off-whitecrystals, 1.222 g). The mother liquor was rotervaped, and the residuewas dissolved in ˜3 ml of EtOAc (with heating), allowed to stand atambient condition for 1 hour, and then 3 mL hexanes was added and storedin a refrigerator for ˜15 hr. A second crop of Example M-5e-1 (greycrystals, 0.133 g) was retrieved similarly. ¹H NMR (400 MHz, DMSO-d₆,δ=2.5 ppm) 12.46 (s, 1H), 3.88 (app br s, 1H), 3.27 (app br s, 1H;overlapped with water signal), 2.28 (br m, 1H), 2.07 (app br s, 1H),1.56 (app s, 1H), 1.40/1.34 (two overlapped s, 9H), 0.71 (m, 1H), 0.45(m, 1H). ¹³C-NMR (100.6 MHz, DMSO-d₆, δ=39.21 ppm) 172.96, 172.60,154.45, 153.68, 78.74, 59.88, 59.58, 36.91, 31.97, 31.17, 27.77, 27.52,14.86, 14.53, 13.69. MP (dec.) for the first crop: 147.5-149.5° C.LC/MS: [M+Na]⁺=250.22. Anal. Calcd. For C₁₁H₁₇NO₄: C, 58.13; H, 7.54; N,6.16. Found (for first crop): C, 58.24; H, 7.84; N, 6.07. Opticalrotation (10 mg/mL in CHCl₃): [α]_(D)=−216 and −212 for the first andsecond crop, respectively.

Example M-5f

Example M-5f was prepared in three steps from Example M-5e-1 accordingto the procedure outlined for the preparation of Example M-1 h from(S)-Boc-proline. [Note: the SEM-regiochemical make up was notdetermined] ¹H NMR (400 MHz, DMSO-d₆, δ=2.5 ppm) 7.46 (d, J=8.6, 2H),7.31 (d, J=8.8, 2H), 6.76 (s, 1H), 5.27 (app br s, 1H), 5.14 (d, J=10.9,1H), 4.60 (app br s, 1H), 3.50-3.37 (m, 3H), 2.24 (m, 2H), 1.80 (app s,12H), 1.61 (br m, 1H), 1.12 (br s, 9H), 0.89-0.73 (m, 2H), 0.70 (br m,1H), 0.56 (br m, 1H), −0.04 (s, 9H). LC/MS (Cond. 2): RT=2.05 min.LC/MS: Anal. Calcd. for [M+H]⁺ C₃₃H₄₉ ⁷⁹BrN₃O₃Si: 642.27. found 642.25.

Example M-5g

Dimethylsulfoxide (1 mL, 14.09 mmol) was added dropwise over 6 min to acooled (−78° C.) dichloromethane (14 mL) solution of oxalyl chloride(0.62 mL, 7.08 mmol), and stirred for 23 min. A dichloromethane (11 mL)solution of Example M-5d (1.0 g, 4.69 mmol; trans/cis ratio=˜25/1) wasadded dropwise to the above mixture over 8 min, and stirring of theheterogeneous mixture was continued at similar temperature for 80 min.Triethylamine (2.5 mL, 17.94 mmol) was added and the reaction wasstirred at −78° C. for 3 hr, then at 0° C. for 1 hr and then the bathwas removed and stirring continued for 10 min. The reaction mixture wasdiluted with dichloromethane (25 mL) and washed with water (15 mL, 2×),dried (MgSO₄) and evaporated in vacuo to afford Example M-5g as a lightbrown oil (1.07 g). The trans/cis stereochemical makeup of the productwas not determined, and it was used in the next step withoutpurification.

Example M-5h

Example M-5h was prepared in two steps from Example M-5g according tothe procedure outlined for the preparation of Example QC-1d from(S)-Boc-prolinal. ¹H NMR (400 MHz, DMSO-d₆, δ=2.5 ppm) 7.14 (app s, 1H),6.81 (d, J=0.8, 1H), 5.57-5.31 (app br s, 1H), 5.22 (d, J=5.22, 1H),4.73 (app br s, 1H), 3.47-3.27 (br m, 3H), 2.24 (app br s, 2H), 1.63(app br s, 1H), 1.45-1.02 (app br m, 9H), 0.89-0.69 (m, 3H), 0.55 (m,1H), −0.04 (s, 9H). LC/MS (Cond. 2): RT=1.60 min. LC/MS: Anal. Calcd.for [M+H]⁺ C₁₉H₃₄N₃O₃Si: 380.24. found 380.21.

Example M-5i

Example M-5i (TFA salt) was prepared in two steps starting from ExampleM-5f and Example M-5h according to the procedure described for thepreparation of Example M-1j from Example M-1 h and Example QC-1d, withthe exception that the crude material was purified with a reverse phaseHPLC (methanol/water/TFA) and then free-based with MCX (6g; MeOH wash;2.0 M NH₃/MeOH elution). Example M-5i was retrieved as an off-white foam(162 mg). ¹H NMR (400 MHz, DMSO-d₆, δ=2.5 ppm) 11.75 (app br s, ˜1H;only one of the imidazole NH was observed), 7.61 (d, J=8.3, 2H), 7.33(app br s, 1H), 7.29 (d, J=8.5, 2H), 6.50 (s, 1H), 3.97 (dd, J=9.8, 7.3,1H), 3.88 (dd, J=10.0, 7.2, 1H), 2.83-2.77 (m, 2H), 2.14-2.03 (m, 2H),1.94-1.76 (m, 14H), 1.44-1.36 (m, 2H), 0.67-0.64 (m, 1H), 0.63-0.60 (m,1H), 0.34-0.27 (m, 2H). LC/MS (Cond. 2): RT=0.94 min. LC/MS: Anal.Calcd. for [M+H]⁺ C₃₀H₃₇N₆: 481.31. found 481.27.

Example M-5

Example M-5 (TFA salt) was prepared from Example M-5i according to theprocedure outlined in Example M-1. ¹H NMR (400 MHz, DMSO-d₆, δ=2.5 ppm):14.13/13.98 (two br s, 2H), 7.99 (br s, 1H), 7.69 (d, J=8.3, 2H), 7.51(d, J=8.3, 2H), 7.30 (s, 1H), 7.25/7.23 (two overlapping d, J=8.5/9.4,1.82H), 6.99-6.93 (br m, 0.18H), 4.98 (m, 1H), 4.90 (m, 1H), 4.40 (m,1.77H), 4.33-4.27 (br m, 0.23H), 3.72 (br m, 2H), 3.54 (s, 3H), 3.53 (s,3H), 2.38-2.07 (m, 4H), 1.96-1.83 (m, 14H), 0.98-0.72 (m, 16H), (Note:the signal of 2H appears to have overlapped with that of the solventsignal). LC/MS (Cond. 2): RT=1.35 min. LC/MS: Anal. Calcd. for [M+H]⁺C₄₄H₅₉N₈O₆: 795.46. found 795.44.

Examples M-6 and M-7

Examples M-6 and M-7 were prepared as TFA salts from Example M-5i andappropriate acids by employing the procedure described for the synthesisof Example M-5, with the exception that an additional purification wasconducted with a second reverse phase HPLC system(water/acetonitrile/TFA) for Example M-7.

RT (LC-Cond.); % homogeneity Example R index; MS data M-6

1.20 min (Cond. 2); >98%; LC/MS: Anal. Calcd. for [M + H]⁺ C₄₂H₅₅N₈O₆:767.42; found 767.47 M-7

1.93 min (Cond. 2a); >95%; LC/MS: Anal. Calcd. for [M + H]⁺ C₅₀H₅₅N₈O₆:863.42; found 863.41

Example M-8

The TFA salt of Example 5 (41 mg) was free based with MCX column (MeOHwash; 2 N NH3/MeOH), and the resultant colorless glassy oil (29 mg) wasdissolved in DMF (2 mL), treated with NCS (10.7 mg, 0.080 mmol) andheated at 50° C. for 15 hr. The reaction mixture was allowed to cool toambient condition, diluted with MeOH and submitted to reverse phase HPLCpurification (XTERRA 30×100 mm S5; water/MeOH/TFA) to afford the TFAsalt of Example 8 as a white foam (27.4 mg). ¹H NMR (400 MHz, DMSO-d₆,δ=2.5 ppm) 7.61 (d, J=8.6, 2H), 7.46 (d, J=8.5, 2H), 7.15 (app t, 2H),4.98-4.92 (m, 2H), 4.43-4.38 (m, 2H), 3.62-3.55 (an overlap of ‘m’ &‘s’, 8H), 2.35-2.21 (m, 4H), 2.10-1.80 (m, 16H), 1.04-0.86 (m, 14H),0.71 (m, 2H). LC/MS (Cond. 4): RT=3.97 min. LC/MS: Anal. Calcd. for[M+H]⁺ C₄₄H₅₈Cl₂N₈O₆: 863.28. found 863.40.

BIOLOGICAL ACTIVITY

An HCV Replicon assay was utilized in the present disclosure, and wasprepared, conducted and validated as described in commonly ownedPCT/US2006/022197 and in O'Boyle et. al. Antimicrob Agents Chemother.2005 April; 49(4):1346-53. Assay methods incorporating luciferasereporters have also been used as described (Apath.com).

HCV-neo replicon cells and replicon cells containing mutations in theNS5A region were used to test the currently described family ofcompounds. The compounds were determined to have more than 10-fold lessinhibitory activity on cells containing mutations than wild-type cells.Thus, the compounds of the present disclosure can be effective ininhibiting the function of the HCV NS5A protein and are understood to beas effective in combinations as previously described in applicationPCT/US2006/022197 and commonly owned WO/04014852. Further, the compoundsof the present disclosure can be effective against the HCV 1b genotype.It should also be understood that the compounds of the presentdisclosure can inhibit multiple genotypes of HCV. Table 2 shows the EC₅₀(Effective 50% inhibitory concentration) values of representativecompounds of the present disclosure against the HCV 1b genotype. In oneembodiment, compounds of the present disclosure are inhibitory versus1a, 1b, 2a, 2b, 3a, 4a, and 5a genotypes. EC₅₀ values against HCV 1b areas follows: A (1-10 μM); B (100-999 nM); C (4.57-99 nM); D (2 nM-4.57nM).

The compounds of the present disclosure may inhibit HCV by mechanisms inaddition to or other than NS5A inhibition. In one embodiment thecompounds of the present disclosure inhibit HCV replicon and in anotherembodiment the compounds of the present disclosure inhibit NS5A.

TABLE 2 1b EC₅₀ Exam- (in nM ple or range) Range Name QC-1 D methyl((1S)-1-(((2S)-2-(4-(4′-(2- ((2S)-1-((2S)-2- ((methoxycarbonyl)amino)-3-methylbutanoyl)-2-pyrrolidinyl)-1H-imidazol-4-yl)-1,1′-bi(3-cyclohexen-1- yl)-4-yl)-1H-imidazol-2-yl)-1-pyrrolidinyl)carbonyl)-2- methylpropyl)carbamate QC-2 C methyl((1S)-2-((2S)-2-(4-(4′-(2-((2S)- 1-(N-(methoxycarbonyl)-L-alanyl)-2-pyrrolidinyl)-1H-imidazol-4-yl)-1,1′- bi(3-cyclohexen-1-yl)-4-yl)-1H-imidazol-2-yl)-1-pyrrolidinyl)-1- methyl-2-oxoethyl)carbamate QC-3 D(1R,1′R)-2,2′-(1,1′-bi(3-cyclohexen-1-yl)-4,4′-diylbis(1H-imidazole-4,2-diyl(2S)-2,1-pyrrolidinediyl))bis(N,N- diethyl-2-oxo-1-phenylethanamine)QC-4 C methyl ((1S)-2-((2S)-2-(4-(4′-(2-((2S)-1-(N-(methoxycarbonyl)-O-methyl-L- seryl)-2-pyrrolidinyl)-1H-imidazol-4-yl)-1,1′-bi(3-cyclohexen-1-yl)-4-yl)-1H-imidazol-2-yl)-1-pyrrolidinyl)-1- (methoxymethyl)-2-oxoethyl)carbamate QC-5 D methyl ((1S,2R)-2-methoxy-1-(((2S)-2-(4-(4′-(2-((2S)-1-(N- (methoxycarbonyl)-O-methyl-L-threonyl)-2-pyrrolidinyl)-1H- imidazol-4-yl)-1,1′-bi(3-cyclohexen-1-yl)-4-yl)-1H-imidazol-2-yl)-1- pyrrolidinyl)carbonyl)propyl)carbamateQC-6 B methyl ((1S)-2-((2S)-2-(4-(4′-(2-((2S)-1-(N-(methoxycarbonyl)-L-alanyl)-2-pyrrolidinyl)-1H-imidazol-4-yl)-1,1′-bi(cyclohexyl)-4-yl)-1H-imidazol-2- yl)-1-pyrrolidinyl)-1-methyl-2-oxoethyl)carbamate QC-7 17.11 A methyl ((1S)-2-((2S)-2-(4-(4′-(2-((2S)-1-(N-(methoxycarbonyl)-O-methyl-L- seryl)-2-pyrrolidinyl)-1H-imidazol-4-yl)-1,1′-bi(cyclohexyl)-4-yl)-1H- imidazol-2-yl)-1-pyrrolidinyl)-1-(methoxymethyl)-2- oxoethyl)carbamate QC-8 2.40 B methyl((1S,2R)-2-methoxy-1-(((2S)- 2-(4-(4′-(2-((2S)-1-(N-(methoxycarbonyl)-O-methyl-L- threonyl)-2-pyrrolidinyl)-1H-imidazol-4-yl)-1,1′-bi(cyclohexyl)-4- yl)-1H-imidazol-2-yl)-1-pyrrolidinyl)carbonyl)propyl)carbamate QC-9 0.011 D methyl((1S)-1-(((2S)-2-(4-(4-(4-(2- ((2S)-1-((2S)-2-((methoxycarbonyl)amino)-3- methylbutanoyl)-2-pyrrolidinyl)-1H-imidazol-4-yl)phenyl)-1-cyclohexen- 1-yl)-1H-imidazol-2-yl)-1-pyrrolidinyl)carbonyl)-2- methylpropyl)carbamate QC-10 C methyl((1S)-2-((2S)-2-(4-(4-(4-(2- ((2S)-1-(N-(methoxycarbonyl)-1-alanyl)-2-pyrrolidinyl)-1H-imidazol-4- yl)-3-cyclohexen-1-yl)phenyl)-1H-imidazol-2-yl)-1-pyrrolidinyl)-1- methyl-2-oxoethyl)carbamate QC-11 Cmethyl ((1S)-2-((2S)-2-(4-(4-(4-(2- ((2S)-1-(N-(methoxycarbonyl)-O-methyl-L-seryl)-2-pyrrolidinyl)-1H- imidazol-4-yl)-3-cyclohexen-1-yl)phenyl)-1H-imidazol-2-yl)-1- pyrrolidinyl)-1-(methoxymethyl)-2-oxoethyl)carbamate QC-12 C methyl ((1S,2R)-2-methoxy-1-(((2S)-2-(4-(4-(4-(2-((2S)-1-(N- (methoxycarbonyl)-O-methyl-L-threonyl)-2-pyrrolidinyl)-1H- imidazol-4-yl)-3-cyclohexen-1-yl)phenyl)-1H-imidazol-2-yl)-1- pyrrolidinyl)carbonyl)propyl)carbamateQC-13 C methyl ((1S)-1-(((2S)-2-(4-(1-(4-(2- ((2S)-1-((2S)-2-((methoxycarbonyl)amino)-3- methylbutanoyl)-2-pyrrolidinyl)-1H-imidazol-4-yl)phenyl)-4-piperidinyl)- 1H-imidazol-2-yl)-1-pyrrolidinyl)carbonyl)-2- methylpropyl)carbamate QC-14 B methyl((1S)-2-((2S)-2-(4-(4-(4-(2- ((2S)-1-(N-(methoxycarbonyl)-O-methyl-L-seryl)-2-pyrrolidinyl)-1H-imidazol-4-yl)-1-piperidinyl)phenyl)-1H-imidazol-2-yl)-1-pyrrolidinyl)-1- (methoxymethyl)-2-oxoethyl)carbamate QC-15 C methyl ((1S)-1-(((2S)-2-(4-(1-(4-(2-((2S)-1-((2S)-2- ((methoxycarbonyl)amino)-3,3-dimethylbutanoyl)-2-pyrrolidinyl)-1H-imidazol-4-yl)phenyl)-4-piperidinyl)- 1H-imidazol-2-yl)-1-pyrrolidinyl)carbonyl)-2,2- dimethylpropyl)carbamate QC-16 D methyl((1S)-1-(((2S)-2-(4-(4-(4-(2- ((2S)-1-((2S)-2-((methoxycarbonyl)amino)-3- methylbutanoyl)pyrrolidin-2-yl)-1H-imidazol-4-yl)phenyl)cyclohexyl)-1H- imidazol-2-yl)pyrrolidin-1-yl)carbonyl)-2- methylpropyl)carbamate (Isomer 1) QC-17 D methyl((1S)-1-(((2S)-2-(4-(4-(4-(2- ((2S)-1-((2S)-2-((methoxycarbonyl)amino)-3- methylbutanoyl)pyrrolidin-2-yl)-1H-imidazol-4-yl)phenyl)cyclohexyl)-1H- imidazol-2-yl)pyrrolidin-1-yl)carbonyl)-2- methylpropyl)carbamate (isomer II) M-1 0.009 D methyl((1S)-1-(((2S)-2-(4-(4-(4-(2- ((2S)-1-((2S)-2-((methoxycarbonyl)amino)-3- methylbutanoyl)-2-pyrrolidinyl)-1H-imidazol-4- yl)phenyl)bicyclo[2.2.2]oct-1-yl)-1H- imidazol-2-yl)-1-pyrrolidinyl)carbonyl)-2- methylpropyl)carbamate M-2 C methyl((1S)-2-((2S)-2-(4-(4-(4-(2- ((2S)-1-(N-(methoxycarbonyl)-L-alanyl)-2-pyrrolidinyl)-1H-imidazol-4-yl)bicyclo[2.2.2]oct-1-yl)phenyl)-1H- imidazol-2-yl)-1-pyrrolidinyl)-1-methyl-2-oxoethyl)carbamate M-3 D methyl ((1R)-2-((2S)-2-(4-(4-(4-(2-((2S)-1-((2R)-2- ((methoxycarbonyl)amino)-2-phenylacetyl)-2-pyrrolidinyl)-1H- imidazol-4-yl)bicyclo[2.2.2]oct-1-yl)phenyl)-1H-imidazol-2-yl)-1- pyrrolidinyl)-2-oxo-1-phenylethyl)carbamate M-4 A methyl ((1S)-1-(((2S)-2-(4-(4-(4-(2-((2S)-1-((2S)-2- ((methoxycarbonyl)amino)-3-methylbutanoyl)-2-pyrrolidinyl)-1H- imidazol-4-yl)phenyl)bicyclo[2.2.2]oct-1-yl)-1,3-oxazol-2-yl)-1-pyrrolidinyl)carbonyl)- 2-methylpropyl)carbamate M-5 Dmethyl ((1S)-1-(((1R,3S,5R)-3-(5-(4- (4-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)amino)-3- methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H- imidazol-5-yl)bicyclo[2.2.2]oct-1-yl)phenyl)-1H-imidazol-2-yl)-2- azabicyclo[3.1.0]hex-2-yl)carbonyl)-2-methylpropyl)carbamate M-6 D methyl ((1S)-1-(((1R,3S,5R)-3-(5-(4-(4-(2-((1R,3S,5R)-2-((2S)-2- ((methoxycarbonyl)amino)butanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H- imidazol-5-yl)bicyclo[2.2.2]oct-1-yl)phenyl)-1H-imidazol-2-yl)-2- azabicyclo[3.1.0]hex-2-yl)carbonyl)propyl)carbamate M-7 D methyl ((1R)-2-((1R,3S,5R)-3-(5-(4-(4-(2-((1R,3S,5R)-2-((2R)-2- ((methoxycarbonyl)amino)-2-phenylacetyl)-2-azabicyclo[3.1.0]hex- 3-yl)-1H-imidazol-5-yl)bicyclo[2.2.2]oct-1-yl)phenyl)-1H- imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)-2-oxo-1- phenylethyl)carbamate M-8 D methyl((1S)-1-(((1R,3S,5R)-3-(4- chloro-5-(4-(4-(4-chloro-2-((1R,3S,5R)-2-((2S)-2- ((methoxycarbonyl)amino)-3- methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H- imidazol-5-yl)bicyclo[2.2.2]oct-1-yl)phenyl)-1H-imidazol-2-yl)-2- azabicyclo[3.1.0]hex-2-yl)carbonyl)-2-methylpropyl)carbamate

The compounds of the present disclosure may inhibit HCV by mechanisms inaddition to or other than NS5A inhibition. In one embodiment thecompounds of the present disclosure inhibit HCV replicon and in anotherembodiment the compounds of the present disclosure inhibit NS5A.

It will be evident to one skilled in the art that the present disclosureis not limited to the foregoing illustrative examples, and that it canbe embodied in other specific forms without departing from the essentialattributes thereof. It is therefore desired that the examples beconsidered in all respects as illustrative and not restrictive,reference being made to the appended claims, rather than to theforegoing examples, and all changes which come within the meaning andrange of equivalency of the claims are therefore intended to be embracedtherein.

What is claimed is:
 1. A compound of Formula (I):

or a pharmaceutically acceptable salt thereof, wherein: s is 0 or 1; Lis -L¹-L²-, wherein L¹ and L² are independently selected from:

 provided that at least one of L¹ and L² is other than

Y and Y′ are independently oxygen (O) or NH; R¹ is hydrogen or—C(O)R^(x); R² is hydrogen or —C(O)R^(y); R^(x) and R^(y) areindependently selected from cycloalkyl, heteroaryl, heterocyclyl,alkoxy, and alkyl substituted with one or more substituentsindependently selected from aryl, alkenyl, cycloalkyl, heterocyclyl,heteroaryl, —OR³, —C(O)OR⁴, —NR^(a)R^(b), and —C(O)NR^(c)R^(d), whereinaryl and heteroaryl may optionally be substituted with one or moresubstituents independently selected from alkyl, haloalkyl, arylalkyl,heterocyclyl, heterocyclylalkyl, halogen, cyano, nitro, —C(O)OR⁴, OR⁵,—NR^(a)R^(b), (NR^(a)R^(b))alkyl, and (MeO)(HO)P(O)O—, and whereincycloalkyl and heterocyclyl may optionally be fused onto an aromaticring and may optionally be substituted with one or more substituentsindependently selected from alkyl, hydroxyl, halogen, aryl,—NR^(a)R^(b), oxo, and —C(O)OR⁴; R³ is hydrogen, alkyl, or arylalkyl; R⁴is alkyl or arylalkyl; R⁵ is hydrogen, alkyl, or arylalkyl; R^(a) andR^(b) are independently selected from hydrogen, alkyl, cycloalkyl,arylalkyl, heteroaryl, —C(O)R⁶, —C(O)OR⁷, —C(O)NR^(c)R^(d), and(NR^(c)R^(d))alkyl, or alternatively, R^(a) and R^(b), together with thenitrogen atom to which they are attached, form a five- or six-memberedring or bridged bicyclic ring structure, wherein said five- orsix-membered ring or bridged bicyclic ring structure optionally maycontain one or two additional heteroatoms independently selected fromnitrogen, oxygen, and sulfur and may contain one, two, or threesubstituents independently selected from C₁ to C₆ alkyl, C₁ to C₄haloalkyl, aryl, hydroxyl, C₁ to C₆ alkoxy, C₁ to C₄ haloalkoxy, andhalogen; R⁶ is alkyl; R⁷ is alkyl, arylalkyl, cycloalkyl, or haloalkyl;R¹⁰⁰ and R¹¹⁰ are independently selected from hydrogen, alkyl,cyanoalkyl, and halo; R^(c) and R^(d) are independently selected fromhydrogen, alkyl, arylalkyl, and cycloalkyl.
 2. The compound according toclaim 1, or a pharmaceutically acceptable salt thereof, wherein L isselected from:


3. The compound according to claim 1, or a pharmaceutically acceptablesalt thereof, wherein Y and Y′ are each NH.
 4. The compound according toclaim 1, or a pharmaceutically acceptable salt thereof, wherein Y isoxygen (O), and Y′ is NH.
 5. The compound of claim 1, or apharmaceutically acceptable salt thereof, wherein: R¹ is —C(O)R^(x); R²is —C(O)R^(y); R^(x) and R^(y) are independently alkyl substituted by atleast one —NR^(a)R^(b), characterized by Formula (A):

wherein: m is 0 or 1; R⁸ is hydrogen or alkyl; R⁹ is selected fromhydrogen, cycloalkyl, aryl, heteroaryl, heterocyclyl, and alkyloptionally substituted with a substituent selected from aryl, alkenyl,cycloalkyl, heterocyclyl, heteroaryl, heterobicyclyl, —OR³, —C(O)OR⁴,—NR^(a)R^(b), and —C(O)NR^(c)R^(d), wherein aryl and heteroaryl mayoptionally be substituted with one or more substituents independentlyselected from alkyl, haloalkyl, arylalkyl, heterocyclyl,heterocyclylalkyl, halogen, cyano, nitro, —C(O)OR⁴, OR⁵, —NR^(a)R^(b),(NR^(a)R^(b))alkyl, and (MeO)(HO)P(O)O—, and wherein cycloalkyl andheterocyclyl may optionally be fused onto an aromatic ring and mayoptionally be substituted with one or more substituents independentlyselected from alkyl, hydroxyl, halogen, aryl, —NR^(a)R^(b), oxo, and—C(O)OR⁴; R¹⁰⁰ and R¹¹⁰ are independently selected from hydrogen andhalo; and R³, R⁴, R⁵, R^(a), R^(b), R^(c), and R^(d) are defined as inclaim
 1. 6. The compound of claim 5, or a pharmaceutically acceptablesalt thereof, wherein: m is 0; R⁸ is hydrogen or C₁ to C₄ alkyl; R⁹ isselected from hydrogen, C₁ to C₆ alkyl optionally substituted with—OR¹², C₃ to C₆ cycloalkyl, allyl, —CH₂C(O)NR^(c)R^(d),(NR^(c)R^(d))alkyl,

wherein j is 0 or 1; k is 1, 2, or 3; n is 0 or an integer selected from1 through 4; each R¹⁰ is independently hydrogen, C₁ to C₄ alkyl, C₁ toC₄ haloalkyl, halogen, nitro, —OBn, or (MeO)(OH)P(O)O—; R¹¹ is hydrogen,C₁ to C₄ alkyl, or benzyl; R¹² is hydrogen, C₁ to C₄ alkyl, or benzyl;R^(a) is hydrogen or C₁ to C₄ alkyl; R^(b) is C₁ to C₄ alkyl, C₃ to C₆cycloalkyl, benzyl, 3-pyridyl, pyrimidin-5-yl, acetyl, —C(O)OR⁷, or—C(O)NR^(c)R^(d); R⁷ is C₁ to C₄ alkyl or C₁ to C₄ haloalkyl; R^(c) ishydrogen or C₁ to C₄ alkyl; and R^(d) is hydrogen, C₁ to C₄ alkyl, or C₃to C₆ cycloalkyl.
 7. The compound of claim 5, or a pharmaceuticallyacceptable salt thereof, wherein: m is 0; R⁸ is hydrogen; R⁹ is phenyloptionally substituted with one up to five substituents independentlyselected from C₁ to C₆ alkyl, C₁ to C₄ haloalkyl, halogen, C₁ to C₆alkoxy, hydroxyl, cyano, and nitro; and NR^(a)R^(b) is a heterocyclyl orheterobicyclyl group selected from:

wherein n is 0, 1, or 2; each R¹³ is independently selected from C₁ toC₆ alkyl, phenyl, trifluoromethyl, halogen, hydroxyl, methoxy, and oxo;and R¹⁴ is C₁ to C₆ alkyl, phenyl, benzyl, or C(O)OR¹⁵ group, whereinR¹⁵ is C₁ to C₄ alkyl, phenyl, or benzyl.
 8. The compound of claim 5, ora pharmaceutically acceptable salt thereof, wherein: m is 1; R⁸ ishydrogen; R⁹ is C₁ to C₆ alkyl, arylalkyl, or heteroarylalkyl; R^(a) ishydrogen; and R^(b) is —C(O)OR⁴, wherein R⁴ is C₁ to C₆ alkyl.
 9. Thecompound of claim 1, or a pharmaceutically acceptable salt thereof,wherein: R¹ is —C(O)R^(x); R² is —C(O)R^(y); and R^(x) and R^(y) areheteroaryl or heterocyclyl independently selected from:

wherein n is 0 or an integer selected from 1 through 4; each R¹³ isindependently selected from hydrogen, C₁ to C₆ alkyl, C₁ to C₄haloalkyl, phenyl, benzyl, C₁ to C₆ alkoxy, C₁ to C₄ haloalkoxy,heterocyclyl, halogen, NR^(c)R^(d), hydroxyl, cyano, and oxo, whereR^(c) and R^(d) are independently hydrogen or C₁ to C₄ alkyl; and R¹⁴ ishydrogen (H), C₁ to C₆ alkyl, benzyl, or —C(O)OR⁴, wherein R⁴ is C₁ toC₆ alkyl.
 10. The compound of claim 1, or a pharmaceutically acceptablesalt thereof, wherein: R¹ is hydrogen or —C(O)R^(x); R² is hydrogen or—C(O)R^(y); R^(x) and R^(y) are cycloalkyl independently selected from:

wherein j is 0, 1, 2, or 3; k is 0, 1, or 2; n is 0 or an integerselected from 1 though 4; each R¹³ is independently selected fromhydrogen, C₁ to C₆ alkyl, C₁ to C₄ haloalkyl, C₁ to C₆ alkoxy, halogen,hydroxyl, cyano, and nitro; and R^(a) and R^(b) are independentlyhydrogen, C₁ to C₆ alkyl, or C(O)OR⁴, wherein R⁴ is C₁ to C₆ alkyl. 11.The compound of claim 1, or a pharmaceutically acceptable salt thereof,wherein: R¹ is —C(O)R^(x); R² is —C(O)R^(y); R^(x) and R^(y) areindependently arylalkyl, wherein aryl part of said arylalkyl mayoptionally be substituted with (NR^(a)R^(b))alkyl; and R^(a) and R^(b)are independently hydrogen, C₁ to C₆ alkyl, or benzyl, or alternatively,R^(a) and R^(b), together with the nitrogen atom to which they areattached, form a five- or six-membered ring selected from

 wherein R¹⁵ is hydrogen, C₁ to C₆ alkyl, or benzyl.
 12. The compound ofclaim 1, or a pharmaceutically acceptable salt thereof, wherein: R¹ is—C(O)R^(x); R² is —C(O)R^(y); and R^(x) and R^(y) are the same and areselected from the group consisting of:

wherein a squiggle bond (

) in the structure indicates that a stereogenic center to which the bondis attached can take either (R)- or (S)-configuration so long aschemical bonding principles are not violated.
 13. The compound of claim1, or a pharmaceutically acceptable salt thereof, wherein: R¹ is—C(O)R^(x); R² is —C(O)R^(y); and R^(x) and R^(y) are both t-butoxy. 14.The compound of claim 1, or a pharmaceutically acceptable salt thereof,wherein R¹ and R² are both hydrogen.
 15. A compound of Formula (II):

or a pharmaceutically acceptable salt thereof, wherein: s is 0 or 1; Lis -L¹-L²-, wherein L¹ and L² are independently selected from:

 provided that at least one of L¹ and L² is other than

Y and Y′ are independently oxygen (O) or NH; R¹ is hydrogen or—C(O)R^(x); R² is hydrogen or —C(O)R^(y); R^(x) and R^(y) areindependently selected from cycloalkyl, heteroaryl, heterocyclyl,alkoxy, and alkyl substituted with one or more substituentsindependently selected from aryl, alkenyl, cycloalkyl, heterocyclyl,heteroaryl, —OR³, —C(O)OR⁴, —NR^(a)R^(b), and —C(O)NR^(c)R^(d), whereinaryl and heteroaryl may optionally be substituted with one or moresubstituents independently selected from alkyl, haloalkyl, arylalkyl,heterocyclyl, heterocyclylalkyl, halogen, cyano, nitro, —C(O)OR⁴, OR⁵,—NR^(a)R^(b), (NR^(a)R^(b))alkyl, and (MeO)(HO)P(O)O—, and whereincycloalkyl and heterocyclyl may optionally be fused onto an aromaticring and may optionally be substituted with one or more substituentsindependently selected from alkyl, hydroxyl, halogen, aryl,—NR^(a)R^(b), oxo, and —C(O)OR⁴; R³ is hydrogen, alkyl, or arylalkyl; R⁴is alkyl or arylalkyl; R⁵ is hydrogen, alkyl, or arylalkyl; R^(a) andR^(b) are independently selected from hydrogen, alkyl, cycloalkyl,arylalkyl, heteroaryl, —C(O)R⁶, —C(O)OR⁷, —C(O)NR^(c)R^(d), and(NR^(c)R^(d))alkyl, or alternatively, R^(a) and R^(b), together with thenitrogen atom to which they are attached, form a five- or six-memberedring or bridged bicyclic ring structure, wherein said five- orsix-membered ring or bridged bicyclic ring structure optionally maycontain one or two additional heteroatoms independently selected fromnitrogen, oxygen, and sulfur and may contain one, two, or threesubstituents independently selected from C₁ to C₆ alkyl, C₁ to C₄haloalkyl, aryl, hydroxyl, C₁ to C₆ alkoxy, C₁ to C₄ haloalkoxy, andhalogen; R⁶ is alkyl; R⁷ is alkyl, arylalkyl, or haloalkyl; and R^(c)and R^(d) are independently selected from hydrogen, alkyl, arylalkyl,and cycloalkyl.
 16. A compound, or a pharmaceutically acceptable saltthereof, selected from the group consisting of:methyl((1S)-1-(((2S)-2-(4-(4′-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amino)-3-methylbutanoyl)-2-pyrrolidinyl)-1H-imidazol-4-yl)-1,1′-bi(3-cyclohexen-1-yl)-4-yl)-1H-imidazol-2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)carbamate;methyl((1S)-2-((2S)-2-(4-(4′-(2-((2S)-1-(N-(methoxycarbonyl)-L-alanyl)-2-pyrrolidinyl)-1H-imidazol-4-yl)-1,1′-bi(3-cyclohexen-1-yl)-4-yl)-1H-imidazol-2-yl)-1-pyrrolidinyl)-1-methyl-2-oxoethyl)carbamate;(1′R)-2,2′-(1,1′-bi(3-cyclohexen-1-yl)-4,4′-diylbis(1H-imidazole-4,2-diyl(2S)-2,1-pyrrolidinediyl))bis(N,N-diethyl-2-oxo-1-phenylethanamine);methyl((1S)-2-((2S)-2-(4-(4′-(2-((2S)-1-(N-(methoxycarbonyl)-O-methyl-L-seryl)-2-pyrrolidinyl)-1H-imidazol-4-yl)-1,1′-bi(3-cyclohexen-1-yl)-4-yl)-1H-imidazol-2-yl)-1-pyrrolidinyl)-1-(methoxymethyl)-2-oxoethyl)carbamate;methyl((1S,2R)-2-methoxy-1-(((2S)-2-(4-(4′-(2-((2S)-1-(N-(methoxycarbonyl)-O-methyl-L-threonyl)-2-pyrrolidinyl)-1H-imidazol-4-yl)-1,1′-bi(3-cyclohexen-1-yl)-4-yl)-1H-imidazol-2-yl)-1-pyrrolidinyl)carbonyl)propyl)carbamate;methyl((1S)-2-((2S)-2-(4-(4′-(2-((2S)-1-(N-(methoxycarbonyl)-L-alanyl)-2-pyrrolidinyl)-1H-imidazol-4-yl)-1,1′-bi(cyclohexyl)-4-yl)-1H-imidazol-2-yl)-1-pyrrolidinyl)-1-methyl-2-oxoethyl)carbamate;methyl((1S)-2-((2S)-2-(4-(4′-(2-((2S)-1-(N-(methoxycarbonyl)-O-methyl-L-seryl)-2-pyrrolidinyl)-1H-imidazol-4-yl)-1,1′-bi(cyclohexyl)-4-yl)-1H-imidazol-2-yl)-1-pyrrolidinyl)-1-(methoxymethyl)-2-oxoethyl)carbamate;methyl((1S,2R)-2-methoxy-1-(((2S)-2-(4-(4′-(2-((2S)-1-(N-(methoxycarbonyl)-O-methyl-L-threonyl)-2-pyrrolidinyl)-1H-imidazol-4-yl)-1,1′-bi(cyclohexyl)-4-yl)-1H-imidazol-2-yl)-1-pyrrolidinyl)carbonyl)propyl)carbamate;methyl((1S)-1-(((2S)-2-(4-(4-(4-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amino)-3-methylbutanoyl)-2-pyrrolidinyl)-1H-imidazol-4-yl)phenyl)-1-cyclohexen-1-yl)-1H-imidazol-2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)carbamate;methyl((1S)-2-((2S)-2-(4-(4-(4-(2-((2S)-1-(N-(methoxycarbonyl)-L-alanyl)-2-pyrrolidinyl)-1H-imidazol-4-yl)-3-cyclohexen-1-yl)phenyl)-1H-imidazol-2-yl)-1-pyrrolidinyl)-1-methyl-2-oxoethyl)carbamate;methyl((1S)-2-((2S)-2-(4-(4-(4-(2-((2S)-1-(N-(methoxycarbonyl)-O-methyl-L-seryl)-2-pyrrolidinyl)-1H-imidazol-4-yl)-3-cyclohexen-1-yl)phenyl)-1H-imidazol-2-yl)-1-pyrrolidinyl)-1-(methoxymethyl)-2-oxoethyl)carbamate;methyl((1S,2R)-2-methoxy-1-(((2S)-2-(4-(4-(4-(2-((2S)-1-(N-(methoxycarbonyl)-O-methyl-L-threonyl)-2-pyrrolidinyl)-1H-imidazol-4-yl)-3-cyclohexen-1-yl)phenyl)-1H-imidazol-2-yl)-1-pyrrolidinyl)carbonyl)propyl)carbamate;methyl((1S)-1-(((2S)-2-(4-(1-(4-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amino)-3-methylbutanoyl)-2-pyrrolidinyl)-1H-imidazol-4-yl)phenyl)-4-piperidinyl)-1H-imidazol-2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)carbamate;methyl((1S)-2-((2S)-2-(4-(4-(4-(2-((2S)-1-(N-(methoxycarbonyl)-O-methyl-L-seryl)-2-pyrrolidinyl)-1H-imidazol-4-yl)-1-piperidinyl)phenyl)-1H-imidazol-2-yl)-1-pyrrolidinyl)-1-(methoxymethyl)-2-oxoethyl)carbamate;methyl((1S)-1-(((2S)-2-(4-(1-(4-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amino)-3,3-dimethylbutanoyl)-2-pyrrolidinyl)-1H-imidazol-4-yl)phenyl)-4-piperidinyl)-1H-imidazol-2-yl)-1-pyrrolidinyl)carbonyl)-2,2-dimethylpropyl)carbamate;methyl((1S)-1-(((2S)-2-(4-(4-(4-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amino)-3-methylbutanoyl)-2-pyrrolidinyl)-1H-imidazol-4-yl)phenyl)bicyclo[2.2.2]oct-1-yl)-1H-imidazol-2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)carbamate;methyl((1S)-2-((2S)-2-(4-(4-(4-(2-((2S)-1-(N-(methoxycarbonyl)-L-alanyl)-2-pyrrolidinyl)-1H-imidazol-4-yl)bicyclo[2.2.2]oct-1-yl)phenyl)-1H-imidazol-2-yl)-1-pyrrolidinyl)-1-methyl-2-oxoethyl)carbamate;methyl((1R)-2-((2S)-2-(4-(4-(4-(2-((2S)-1-((2R)-2-((methoxycarbonyl)amino)-2-phenylacetyl)-2-pyrrolidinyl)-1H-imidazol-4-yl)bicyclo[2.2.2]oct-1-yl)phenyl)-1H-imidazol-2-yl)-1-pyrrolidinyl)-2-oxo-1-phenylethyl)carbamate;methyl((1S)-1-(((2S)-2-(4-(4-(4-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amino)-3-methylbutanoyl)-2-pyrrolidinyl)-1H-imidazol-4-yl)phenyl)bicyclo[2.2.2]oct-1-yl)-1,3-oxazol-2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)carbamate;methyl((1S)-1-(((1R,3S,5R)-3-(5-(4-(4-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-5-yl)bicyclo[2.2.2]oct-1-yl)phenyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)carbonyl)-2-methylpropyl)carbamate;methyl((1S)-1-(((1R,3S,5R)-3-(5-(4-(4-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)amino)butanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-5-yl)bicyclo[2.2.2]oct-1-yl)phenyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)carbonyl)propyl)carbamate;methyl((1R)-2-((1R,3S,5R)-3-(5-(4-(4-(2-(1R,3S,5R)-2-((2R)-2-((methoxycarbonyl)amino)-2-phenylacetyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-5-yl)bicyclo[2.2.2]oct-1-yl)phenyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)-2-oxo-1-phenylethyl)carbamate;methyl((1S)-1-(((2S)-2-(4-(4-(4-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amino)-3-methylbutanoyl)pyrrolidin-2-yl)-1H-imidazol-4-yl)phenyl)cyclohexyl)-1H-imidazol-2-yl)pyrrolidin-1-yl)carbonyl)-2-methylpropyl)carbamate;methyl((1S)-1-(((1R,3S,5R)-3-(4-chloro-5-(4-(4-(4-chloro-2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-5-yl)bicyclo[2.2.2]oct-1-yl)phenyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)carbonyl)-2-methylpropyl)carbamate;and corresponding stereoisomers thereof.
 17. A compound which is


18. A compound which is


19. A composition comprising a compound of claim 1, or apharmaceutically acceptable salt thereof, and a pharmaceuticallyacceptable carrier.
 20. A method of treating an HCV infection in apatient, comprising administering to the patient a therapeuticallyeffective amount of a compound of claim 1, or a pharmaceuticallyacceptable salt thereof.